2-(substituted pyrrolidinylthio)carbapenem derivatives

ABSTRACT

A compound of the formula:    &lt;IMAGE&gt;  (I)  wherein R is a hydrogen atom or a methyl group, R1 is a hydrogen atom or a negative charge, each of R2 and R3 which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, a formimidoyl group, an acetoimidoyl group, -COOR4, -CON(R5)R6, -N(R5)R6, -CH2COOR4, -CH2N(R5)R6 or -CH2CON(R5)R6 (wherein R4 is a hydrogen atom or a lower alkyl group, each of R5 and R6 which may be the same or different, is a hydrogen atom or a lower alkyl group, or R5 and R6 form together with the adjacent nitrogen atom a heterocyclic group selected from the group consisting of an aziridinyl group, an azetidinyl group, a pyrrolidinyl group and a piperidyl group), A is =NR7, =N+(R7)R8, wherein each of R7 and R8 which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, a formimidoyl group, an acetoimidoyl group, -COOR4, -CON(R5)R6, -N(R5)R6, -CH2COOR4, -CH2N(R5)R6 or -CH2CON(R5)R6 (wherein R4, R5 and R6 are as defined above), p is an integer of from 0 to 3, and q is an integer of from 1 to 3; or a pharmaceutically acceptable salt or ester thereof.

This application is a continuation-in-part of U.S. application Ser. No. 07/674,971, filed Mar. 26, 1991, abandoned.

The present invention relates to novel carbapenem (7-oxo-1-azabicyclo[3.2.0]hept-2-en-2-carboxylic acid) compounds, and antibacterial agents containing such compounds as active ingredients, and a process for producing such compounds.

In recent years, new β-lactam antibiotic substances have been found in nature which have the same β-lactam rings as penicillin derivatives and as cephalosporin derivatives, but which have different basic structures.

For example, naturally derived carbapenem compounds such as thienamycin isolated from the fermentation of Streptomyces cattleya (J. Am. Chem. Soc., vol. 100, p.6491 (1978)), may be mentioned. Thienamycin has an excellent antibacterial spectrum and strong antibacterial activities over a wide range against gram positive bacteria and gram negative bacteria. Therefore, its development as a highly useful β-lactam agent has been expected. However, thienamycin itself is chemically unstable, and it has been reported that it is likely to be decomposed by a certain enzyme in vivo such as renal dehydropeptidase I (hereinafter referred to simply as DHP-I), whereby the antibacterial activities tend to decrease, and the recovery rate in the urine is low (Antimicrob. Agents Chemother., vol. 22, p.62 (1982); ditto, vol. 23, p.300 (1983)).

Merck & Co., Inc. have synthesized many thienamycin analogues with an aim to maintain the excellent antibacterial activities of thienamycin and to secure chemical stability. As a result, imipenem, (5R,6S,8R)-3-[[2-(formimidoylamino)ethyl]thio]-6-(1-hydroxyethyl)-7-oxo-1-azabicyclo[3.2.0]hept-2-en-2-carboxylic acid monohydrate, obtained by formimidation of the amine group of thienamycin, has been practically developed as a pharmaceutical product (J. Med. Chem., vol. 22, p. 1435 (1979)). Imipenem has antibacterial activities of an equal or higher level than thienamycin against various types of bacteria and has β-lactamase resistance. Especially against Pseudomonas aeruqinosa, its antibacterial activities are superior to thienamycin by from 2 to 4 times. Further, the stability of imipenem in the solid form or in an aqueous solution is remarkably improved over thienamycin.

However, like thienamycin, imipenem is likely to be decomposed by DHP-I in the human kidney. Therefore, it can not be used for treatment of the urinary-tract infection. Further, it presents toxicity against the kidney due to the decomposition products. Therefore, imipenem can not be administered alone and is required to be used in combination with a DHP-I inhibitor like cilastatin (Antimicrob. Agents Chemother., vol. 12 (Suppl. D), p. 1 (1983)). In recent years, imipenem has been frequently used for the treatment and prevention of infectious diseases. Consequently, highly methicillin resistant Staphylococcus aureus which is resistant to imipenem and imipenem resistant Pseudomonas aeruqinosa are increasing in the clinical field. Imipenem does not show adequate treating effects against these resistant bacteria.

As the prior art closest to the present invention, U.S. Pat. No. 4,933,333 may be mentioned. This publication discloses carbapenem compounds having a 2-(aminocarbonyl or N-mono- or N,N-di-lower alkylaminocarbonyl)pyrrolidin-4-ylthio group at the 2-position of the carbapenem structure, represented by meropenem, SM-7338: (4R,5S,6S,8R,2'S,4'S)-6-(1-hydroxyethyl)-4-methyl-3-[2-(N,N-dimethylaminocarbonyl)-pyrrolidin-4-ylthio]-7-oxo-1-azabicyclo[3.2.0]hept-2-en-2-carboxylic acid, as a typical compound.

β-Lactam antibiotics exhibit selective toxicity against bacteria and show no substantial effects against animal cells. Therefore, they are widely used for treatment of infectious diseases caused by bacteria, as rare antibiotics having little side effects, and thus are highly useful drugs.

However, in recent years, highly methicillin resistant Staphylococcus aureus and resistant Pseudomonas aeruginosa have been isolated frequently from patients with the immunity decreased, as bacteria causing hardly curable infectious diseases. This is regarded as a clinically serious problem. Accordingly, it is strongly desired to develop an antibacterial agent having improved antibacterial activities against such resistant bacteria. Especially with respect to carbapenem compounds, it is desired to improve the antibacterial activities, to improve the stability against DHP-I, to reduce the toxicity against the kidney and to reduce side effects against the central nervous system.

The compounds disclosed in U.S. Pat. No. 4,933,333, particularly meropenem, have the stability against DHP-I substantially improved. However, the antibacterial activities against the above-mentioned highly methicillin resistant Staphylococcus aureus are not adequate, and carbapenem compound having superior antibacterial activities, is desired.

The present inventors have made extensive researches with an aim to provide novel carbapenem compounds which have excellent antibacterial activities and which are resistant against DHP-I. As a result, they have found that carbapenem compounds of the present invention having, at the 2-position of the carbapenem structure, group of the formula: ##STR2## wherein each of R² and R³ which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, a formimidoyl group, an acetoimidoyl group, --COOR⁴, --CON(R⁵)R⁶, --N(R⁵)R⁶, --CH₂ COOR⁴, --CH₂ N(R⁵)R⁶ or --CH₂ CON(R⁵)R⁶ (wherein R⁴ is a hydrogen atom or a lower alkyl group, each of R⁵ and R⁶ which may be the same or different, is a hydrogen atom or a lower alkyl group, or R⁵ and R⁶ form together with the adjacent nitrogen atom a heterocyclic group selected from the group consisting of an aziridinyl group, an azetidinyl group, a pyrrolidinyl group and a piperidyl group), A is ═NR⁷, ═N⁺ (R⁷)R⁸, --CON(R⁷)--, --CON(R⁷)CO--, --CON(R⁷)CON(R⁸)--, --N(R⁷)CO(CH₂)_(s) N(R⁸)--, --N(R⁷)CO(CH₂ )_(s) CON(R⁵)--, --CON(R⁷)N(R⁸)-- or --N(R⁷)(CH₂)_(s) N(R⁸)-- {wherein each of R⁷ and R⁸ which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, a formimidoyl group, an acetoimidoyl group, 13 COOR⁴, --CON(R⁵)R⁶, --N(R⁵)R⁶, --CH₂ COOR⁴, --CH₂ N(R⁵)R⁶ or --CH₂ CON(R⁵)R⁶ (wherein R⁴, R⁵ and R⁶ are as defined above), s is an integer of from 1 to 3}, p is an integer of from 0 to 3, and each of q and r which may be the same or different, is an integer of from 0 to 5, provided that q and r are not simultaneously 0 and q+r≦6, are novel compounds not disclosed any literatures, and that such compounds have strong antibacterial activities against gram positive bacteria such as Staphylococcus aureus and against gram negative bacteria including Pseudomonas aeruginosa and further exhibit excellent stability against DHP-I. The present invention has been accomplished on the basis of this discovery.

The present invention provides a compound of the formula: ##STR3## wherein R is a hydrogen atom or a methyl group, R¹ is a hydrogen atom or a negative charge, each of R² and R³ which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, a formimidoyl group, an acetoimidoyl group, --COOR⁴, --CON(R⁵)R⁶, --N(R⁵)R⁶, --CH₂ COOR⁴, --CH₂ N(R⁵)R⁶ or --CH₂ CON(R⁵)R⁶ (wherein R⁴ is a hydrogen atom or a lower alkyl group, each of R⁵ and R⁶ which may be the same or different, is a hydrogen atom or a lower alkyl group, or R⁵ and R⁶ form together with the adjacent nitrogen atom a heterocyclic group selected from the group consisting of an aziridinyl group, an azetidinyl group, a pyrrolidinyl group and a piperidyl group), A is ═NR⁷, ═N⁺ (R⁷)R⁸, --CON(R⁷)--, --CON(R⁷)CO--, --CON(R⁷)CON(R⁵)--, --N(R⁷)CO(CH₂)_(s) N(R⁸)-, --N(R⁷)CO(CH₂)_(s) CON(R⁵)--, --CON(R⁷)N(R₈)-- or --N(R⁷)(CH₂)_(s) N(R⁵)-- {wherein each of R⁷ and R⁸ which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, a formimidoyl group, an acetoimidoyl group, --COOR⁴, --CON(R⁵)R⁶, --N(R⁵)R⁶, --CH₂ COOR⁴, --CH₂ N(R⁵)R⁶ or --CH₂ CON(R₅)R⁶ (wherein R⁴, R⁵ and R⁶ are as defined above), s is an integer of from 1 to 3}, p is an integer of from 0 to 3, and each of q and r which may be the same or different, is an integer of from 0 to 5, provided that q and r are not simultaneously 0 and q+r≦6; or a pharmaceutically acceptable salt or ester thereof.

The present invention also provides a process for producing the compound of the formula (I) or a pharmaceutically acceptable salt or ester thereof, which comprises reacting a compound of the formula: ##STR4## wherein R is as defined above, R¹³ is a hydrogen atom or a hydroxyl-protecting group, and R¹⁴ is a hydrogen atom or a carboxyl-protecting group, or a reactive derivative thereof, with a compound of the formula: ##STR5## wherein R¹⁵ is a hydrogen atom or an imino-protecting group, each of R²⁰ and R³⁰ which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group which may be protected, a formimidoyl group which may be protected, an acetoimidoyl group which may be protected, --COOR⁴⁰ --CON(R⁵⁰ R⁶⁰) --N(R⁵⁰)R⁶⁰ --CH₂ COOR⁴⁰, --CH₂ N(R⁵⁰)R⁶⁰ or --CH₂ CON(R⁵⁰)R⁶⁰ (wherein R⁴⁰ is a hydrogen atom, a lower alkyl group or a carboxyl-protecting group, and each of R⁵⁰ and R⁶⁰ which may be the same or different, is a hydrogen atom, a lower alkyl group, an amino-protecting group or an imino-protecting group, or R⁵⁰ and R⁶⁰ form together with the adjacent nitrogen atom a heterocyclic group selected from the group consisting of an aziridinyl group, an azetidinyl group, a pyrrolidinyl group and a piperidyl group), B is ═NR⁷⁰, ═N⁺ (R⁷⁰)R⁸⁰ , --CON(R⁷⁰)--, --CON(R⁷⁰)CO--, --CON(R⁷⁰)CON(R⁸⁰)--, --N(R⁷⁰)CO(CH₂)_(s) N(R⁸⁰ (--, --N(R⁷⁰)CO(CH₂)_(s) CON(R⁸⁰)--, --CON(R⁷⁰)N(R⁸⁰)-- or --N(R⁷⁰)(CH₂)_(s) N(R⁸⁰)-- {wherein each of R⁷⁰ and R⁸⁰ which may be the same or different is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group which may be protected, a formimidoyl group which may be protected, an acetoimidoyl group which may be protected, an imino-protecting group, --COOR⁴⁰, --CON(R⁵⁰)R⁶⁰, --N(R⁵⁰)R⁶⁰, --CH₂ COOR⁴⁰, --CH₂ N(R⁵⁰)R⁶⁰ or --CH₂ CON(R⁵⁰)R⁶⁰ (wherein R⁴⁰, R⁵⁰ and R⁶⁰ are as defined above), and s is an integer of from 1 to 3}, and p, q and r are as defined above, to obtain a compound of the formula: ##STR6## wherein R, R¹³, R¹⁴, R¹⁵, R²⁰, R³⁰, B p, q and r are as defined above, and if necessary, removing any protecting group of the compound of the formula (IV).

Further, the present invention provides an antibacterial agent comprising an antibacterially effective amount of the compound of the-formula (I) or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier or diluent.

Now, the present invention will be described in detail with reference to the preferred embodiments. Firstly, the symbols and terms used in this specification will be explained.

The compound of the present invention has a basic structure of the formula: ##STR7## which is systematically referred to as a 7-oxo-1-azabicyclo[3.2.0]hept-2-en-2-carboxylic acid. For the convenience sake, in this specification, this basic structure will be referred to as a 1-carbapen-2-em-3-carboxylic acid by putting the numbers based on a commonly widely used carbapenem of the formula: ##STR8##

The present invention includes optical isomers based on the asymmetrical carbon atoms at the 1-position, 5-position, 6-position and 8-position of the carbapenem structure and stereoisomers. Among these isomers, preferred is a compound of a (5R,6S,8R) configuration i.e. a compound having a steric configuration of (5R,6S) (5,6-trans) like thienamycin and in which the carbon atom at the 8-position takes a R-configuration, or a compound of a (1R,5S,6S,8R) configuration in a case where a methyl group is present at the 1-position.

Also with respect to the 2-(allcyclic heteroring-substituted or alicyclic heteroring lower alkyl)pyrrolidin-4-ylthio group, the present invention includes isomers based on the asymmetrical carbon atoms at the 2-position and 4-position of the pyrrolidine structure and in the side chain at the 2-position. Among these isomers, preferred are compounds of (2'S,4'S) configuration and (2'R,4'R) configuration when p is 0, and compounds of (2'R,4'S) configuration and (2'S,4'R) configuration when p is an integer of from 1 to 3.

Further, with respect to the alicyclic heterocyclic group at the 2-position of the pyrrolidine structure, there exist isomers based on asymmetrical carbons, and the present invention includes such isomers as well.

The lower alkyl group means a linear or branched alkyl group having from 1 to 6 carbon atoms, such as a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, a sec-butyl group, a tert-butyl group, a pentyl group or a hexyl group, preferably a methyl group, an ethyl group or a tert-butyl group.

The hydroxy lower alkyl group means a hydroxyalkyl group having the above mentioned lower alkyl group substituted with a hydroxyl group, such as a hydroxymethyl group, a hydroxyethyl group, a hydroxypropyl group or a hydroxybutyl group, preferably a hydroxymethyl group or a hydroxyethyl group.

The lower alkyl carbamoyl group means a carbamoyl group substituted by the above mentioned lower alkyl group, such as an N-methylcarbamoyl group, an N-ethylcarbamoyl group or an N-propylcarbamoyl group, preferably an N-methylcarbamoyl group.

The di-lower alkyl carbamoyl group means a carbamoyl group di-substituted by the above mentioned lower alkyl group, such as an N,N-dimethyl carbamoyl group, an N,N-diethyl carbamoyl group or an N-ethyl-N-methylcarbamoyl group, preferably an N,N-dimethylcarbamoyl group.

The carboxyl-protecting group may, for example, be a lower alkyl group such as a methyl group, an ethyl group, a propyl group, an isopropyl group or a tert-butyl group; a halogenated lower alkyl group such as a 2,2,2-trichloroethyl group or a 2,2,2-trifluoroethyl group; a lower alkanoyloxyalkyl group such as an acetoxymethyl group, a propionyloxymethyl group, a pivaloyloxymethyl group, a 1-acetoxyethyl group-or a 1-propionyloxyethyl group; a lower alkoxycarbonyloxyalkyl group such as a 1-(methoxycarbonyloxy)ethyl group, a 1-(ethoxycarbonyloxy)ethyl group or a 1-(isopropoxycarbonyloxy)ethyl group; a lower alkenyl group such as a 2-propenyl group, a 2-chloro-2-propenyl group, a 3-methoxycarbonyl-2-propenyl group, a 2-methyl-2-propenyl group, a 2-butenyl group or a cinnamyl group; an aralkyl group such as a benzyl group, a p-methoxybenzyl group, a 3,4-dimethoxybenzyl group, an o-nitrobenzyl group, a p-nitrobenzyl group, a benzhydryl group or a bis(p-methoxyphenyl)methyl group; a (5-substituted 2-oxo-1,3-dioxol-4-yl)methyl group such as a (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl group; a lower alkylsilyl group such as a trimethylsilyl group or a tert-butyldimethylsilyl group; an indanyl group, a phthalidyl group or a methoxymethyl group. Particularly preferred are a 2-propenyl group, a p-nitrobenzyl group, a p-methoxybenzyl group, a benzhydryl group and a tert-butyldimethylsilyl group.

The hydroxyl-protecting group may, for example, be a lower alkylsilyl group such as a trimethylsilyl group or a tert-butyldimethylsilyl group; a lower alkoxymethyl group such as a methoxymethyl group or a 2-methoxyethoxymethyl group; a tetrahydropyranyl group; an aralkyl group such as a benzyl group, a p-methoxybenzyl group, a 2,4-dimethoxybenzyl group, an o-nitrobenzyl group, a p-nitrobenzyl group or a trityl group; an acyl group such as a formyl group or an acetyl group; a power alkoxycarbonyl group such as a tert-butoxycarbonyl group, a 2-iodoethoxycarbonyl group or a 2,2,2-trichloroethoxycarbonyl group; an alkenyloxycarbonyl group such as a 2-propenyloxycarbonyl group, a 2-chloro-2-propenyloxycarbonyl group, a 3-methoxycarbonyl-2-propenyloxycarbonyl group, a 2-methyl-2-propenyloxycarbonyl group, a 2-butenyloxycarbonyl group or a cinnamyloxycarbonyl group; or an aralkyloxycarbonyl group such as a benzyloxycarbonyl group, a p-methoxybenzyloxycarbonyl group, an o-nitrobenzyloxycarbonyl group or a p-nitrobenzyloxycarbonyl group. Particularly preferred are a 2-propenyloxycarbonyl group, a p-nitrobenzyloxycarbonyl group and a tert-butyldimethylsilyl group.

The amino- or imino-protecting group may, for example, be an aralkylidene group such as a benzylidene group, a p-chlorobenzylidene group, a p-nitrobenzylidene group, a salicylidene group, an α-naphthylidene group or a β-naphthylidene group; an aralkyl group such as a benzyl group, a p-methoxybenzyl group, a 3,4-dimethoxybenzyl group, an o-nitrobenzyl group, a p-nitrobenzyl group, a benzhydryl group, a bis(p-methoxyphenyl)methyl group or a trityl group; a lower alkanoyl group such as a formyl group, an acetyl group, a propionyl group, a butyryl group, an oxalyl group, a succinyl group, or a pivaloyl-group; a halogenated lower alkanoyl group such as a chloroacetyl group, a dichloroacetyl group, a trichloroacetyl group or a trifluoroacetyl group; an arylalkanoyl group such as a phenylacetyl group or a phenoxyacetyl group; a lower alkoxycarbonyl group such as a methoxycarbonyl group, an ethoxycarbonyl group, a propoxycarbonyl group or a tert-butoxycarbonyl group; a halogenated lower alkoxycarbonyl group such as a 2-iodoethoxycarbonyl group or a 2,2,2-trichloroethoxycarbonyl group; an alkenyloxycarbonyl group such as a 2-propenyloxycarbonyl group, a 2-chloro-2-propenyloxycarbonyl group, a 3-methoxycarbonyl-2-propenyloxycarbonyl group, a 2-methyl-2-propenyloxycarbonyl group, a 2-butenyloxycarbonyl group or a cinnamyloxycarbonyl group; an aralkyloxycarbonyl group such as a benzyloxycarbonyl group, an o-nitrobenzyloxycarbonyl group, a p-nitrobenzyloxycarbonyl group or a phenethyloxycarbonyl group; or a lower alkylsilyl group such as a trimethylsilyl group or a tert-butyldimethylsilyl group. Particularly preferred are a 2-propenyloxycarbonyl group, a tert-butoxycarbonyl group and a p-nitrobenzyloxycarbonyl group.

The alicyclic heterocyclic group on the pyrrolidin-4-ylthio group as the side chain at the 2-position of the carbapenem structure, is substituted at the 2-position of the pyrrolidine ring and has a structure of the formula: ##STR9## wherein each of R² and R³ which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, a formimidoyl group, an acetoimidoyl group, --COOR⁴, --CON(R⁵)R⁶, --N(R⁵)R⁶, --CH₂ COOR⁴, --CH₂ N(R⁵)R⁶ or --CH₂ CON(R⁵)R⁶ (wherein R⁴ is a hydrogen atom or a lower alkyl group, each of R⁵ and R⁶ which may be the same or different, is a hydrogen atom or a lower alkyl group, or R⁵ and R⁶ form together with the adjacent nitrogen atom a heterocyclic group selected from the group consisting of an aziridinyl group, an azetidinyl group, a pyrrolidinyl group and a piperidyl group), A is ═NR⁷, ═N⁺ (R⁷)R⁸ --CON (R⁷)--, --CON(R⁷)CO--, --CON(R⁷)CON(R⁸)--, --N(R⁷)CO(CH₂)_(s) N(R⁸)--, --N(R⁷)CO(CH₂)_(s) CON(R⁸)--, --CON(R⁷)N(R⁸)-- or --N(R⁷)(CH₂)_(s) N(R⁸)-- {wherein each of R⁷ and R⁸ which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, a formimidoyl group, an acetoimidoyl group, --COOR⁴, --CON(R⁵)R⁶, --N(R⁵)R⁶, --CH₂ COOR⁴, --CH₂ N(R⁵)R⁶ or --CH₂ CON(R⁵)R⁶ (wherein R⁴, R⁵ and R⁶ are as defined above), s is an integer of from 1 to 3}, p is an integer of from 0 to 3, and each of q and r which may be the same or different, is an integer of from 0 to 5, provided that q and r are not simultaneously 0 and q+r≦6. R² (or R²⁰) and R³ (or R³⁰) may be the same or different and may be substituted at any optional positions on carbon atoms constituting said alicyclic heteroring. Each of R² and R³ is preferably a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, --CON(R⁵)R⁶ or --N(R⁵)R⁶ (wherein R⁵ and R⁶ are as defined above). Particularly preferred among them is a hydrogen atom, a carbamoyl group, a lower alkyl carbamoyl group, a di-lower alkyl carbamoyl group or an amino-group.

A represents a partial Structure of said alicyclic heterocyclic group.

A is preferably ═NR⁷, ═N⁺ (R⁷)R^(s), --CON(R⁷)--, --CON(R⁷)CO--, --CON(R⁷)CON(R⁸)--, --N(R⁷)COCH₂ N(R⁸)--, --CON(R⁷)N(R⁵)-- or --N(R⁷)(CH₂)₂ N(R⁸)-- (wherein R⁷ and R⁸ are as defined above). Among them, ═NR⁷, ═N⁺ (R⁷)R⁸ or --CON(R⁷)-- (wherein R⁷ and R⁸ are as defined above) is preferable. Particularly preferred among them is ═NH, ═NMe, ═N⁺ Me₂ or --CONH--.

p is an integer of from 0 to 3, preferably 0 or 1, more preferably 0.

When p is 0, said alicyclic heterocyclic group may be a substituent selected from the group consisting of an aziridinyl group, an azetidinyl group, a 2-carbamoylazetidinyl group, a 2-oxoazetidinyl group, an N-methyl-2-oxoazetidinyl group, a pyrrolidinyl group, an N-methylpyrrolidinyl group, an N,N-dimethylpyrrolidinio group, a 2-oxopyrrolidinyl group, a 2,5-dioxopyrrolidinyl group, an N-(2-hydroxyethyl)pyrrolidinyl group, a 2,5-dioxo-N-methylpyrrolidinyl group, a 2-carbamoylpyrrolidinyl group, a 2-(N-methylcarbamoyl)pyrrolidinyl group, a 2-(N,N-dimethylcarbamoyl)pyrrolidinyl group, a 3-amino-2-oxopyrrolidinyl group, a pyrazolidinyl group, a 3-oxopyrazolidinyl group, an imidazolidinyl group, a 2,4-dioxoimidazolidinyl group, a piperazinyl group, a 2-oxopiperazinyl group, a piperidyl group, an N-methylpiperidyl group, an N,N-dimethylpiperidinic group, a 2-oxopiperidyl group, a 2,6-dioxopiperidyl group, a 2-carbamoyl piperidyl group, a hexahydroazepinyl group, an N-methylhexahydroazepinyl group, an N,N-dimethylhexahydroazepinio group, a hexahydro-2-oxoazepinyl group, a 2,7-dioxohexahydroazepinyl group, a 2-carbamoylhexahydroazepinyl group, a hexahydro-1H-1,4-diazepinyl group, a hexahydro-2-oxo-1H-1,4-diazepinyl group, an octahydroazocinyl group, an N-methyloctahydroazocinyl group and an N,N-dimethyloctahydroazocinio group. Particularly preferred among them is a substituent selected from the group consisting of a 2-oxoazetidinyl group, a pyrrolidinyl group, an N,N-dimethylpyrrolidinio group, a 2-carbamoylpyrrolidinyl group, a 3-amino-2-oxopyrrolidinyl group, a 2-oxopyrrolidinyl group, a piperidyl group and a 2-oxopiperidyl group.

When p is 1, it may be a substituent selected from the group consisting of an aziridinylmethyl group, an azetidinylmethyl group, a 2-carbamoylazetidinylmethyl group, a 2-oxoazetidinylmethyl group, an N-methyl-2-oxoazetidinylmethyl group, a pyrrolidinylmethyl group, an N-methylpyrrolidinylmethyl group, an N,N-dimethylpyrrolidiniomethyl group, a 2-oxopyrrolidinylmethyl group, a 2,5-dioxopyrrolidinylmethyl group, an N-(2-hydroxyethyl)pyrrolidinylmethyl group, a 2,5-dioxo-N-methylpyrrolidinylmethyl group, a 2-carbamoylpyrrolidinylmethyl group, a 2-(N-methylcarbamoyl)pyrrolidinylmethyl group, a 2-(N,N-dimethylcarbamoyl)pyrrolidinylmethyl group, a 3-amino-2-oxopyrrolidinylmethyl group, a pyrazolidinylmethyl group, a 3-oxopyrazolidinylmethyl group, an imidazolidinylmethyl group, a 2,4-dioxoimidazolidinylmethyl group, a piperazinylmethyl group, a 2-oxopiperazinylmethyl group, a piperidylmethyl group, an N-methylpiperidylmethyl group, an N,N-dimethylpiperidiniomethyl group, a 2-oxopiperidylmethyl group, a 2,6-dioxopiperidylmethyl group, a 2-carbamoylpiperidylmethyl group, a hexahydroazepinylmethyl group, an N-methylhexahydroazepinylmethyl group, an N,N-dimethylhexahydroazepiniomethyl group, a hexahydro-2-oxoazepinylmethyl group, a 2,7-dioxohexahydroazepinylmethyl group, a 2-carbamoylhexahydroazepinylmethyl group, a hexahydro-1H-1,4-diazepinylmethyl group, a hexahydro-2-oxo-1H-1,4-diazepinylmethyl group, an octahydroazocinylmethyl group, an N-methyloctahydroazocinylmethyl group and an N,N-dimethyloctahydroazociniomethyl group. Particularly preferred among them is a substituent selected from the group consisting of a 2-oxoazetidinylmethyl group, a pyrrolidinylmethyl group, an N,N-dimethylpyrrolidiniomethyl group, a 2-carbamoylpyrrolidinylmethyl group, a 3-amino-2-oxopyrrolidinylmethyl group, a 2-oxopyrrolidinylmethyl group, a piperidylmethyl group and a 2-oxopiperidylmethyl group.

R¹ is a hydrogen atom or a negative charge. When the alicyclic heterocyclic group substituted at the 2-position of the pyrrolidine ring has a quaternary ammonium structure, R¹ represents a negative charge forming a pair with the ammonium ion, whereby the compound of the formula (I) forms an intramolecular salt.

The salt of the compound of the formula (I) is a common pharmaceutically acceptable salt and may, for example, be a salt at the carboxyl group at the 3-position of the carbapenem structure, or at the pyrrolidine base or the base of the alicyclic heterocyclic group in the side chain at the 2-position of the carbapenem structure.

The basic addition salt at said carboxyl group includes, for example, an alkali metal salt such as a sodium salt or a potassium salt; an alkaline earth metal salt such as a calcium salt or a magnesium salt; an ammonium salt; an aliphatic amine salt such as a trimethylamine salt, a triethylamine salt, a dicyclohexylamine salt, an ethanolamine salt, a diethanolamine salt, a triethanolamine salt or a procaine salt; an aralkylamine salt such as an N,N'-dibenzylethylenediamine salt; an aromatic heterocyclic amine salt such as a pyridine salt, a picoline salt, a quinoline salt or an isoquinoline salt; a quaternary ammonium salt such as a tetramethylammonium salt, a tetraethylammonium salt, a benzyltrimethylammonium salt, a benzyltriethylammonium salt, a benzyltributylammonium salt, a methyltrioctylammonium salt or a tetrabutylammonium salt; and a basic amino acid salt such as an arginine salt or a lysine salt.

The acid addition salt at the pyrrolidine base or at the base of the alicyclic heterocyclic group includes, for example, an inorganic salt such as a hydrochloride, a sulfate, a nitrate, a phosphate, a carbonate, a hydrogencarbonate or a perchlorate; an organic salt such as an acetate, a propionate, a lactate, a maleate, a fumarate, a tartrate, a malate, a succinate or an ascorbate; a sulfonate such as a methanesulfonate, an isethionate, a benzenesulfonate or a p-toluenesulfonate; and an acidic amino acid salt such as an aspartate or a glutamate.

The non-toxic ester of the compound of the formula (I) means a common pharmaceutically acceptable ester at the carboxyl group at the 3-position of the carbapenem structure. For example, it includes an ester with an alkanoyloxymethyl group such as an acetoxymethyl group or a pivaloyloxymethyl group, an ester with an alkoxycarbonyloxyalkyl group such as a 1-(ethoxycarbonyloxy)ethyl group, an ester with a phthalidyl group and an ester with a (5-substituted-2-oxo-1,3-dioxol-4-yl)methyl group such as a (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl group.

The compound of the formula (I) of the present invention includes a compound of the formula: ##STR10## wherein R is a hydrogen atom or a methyl group, A¹ is ═NR⁹, --CON(R¹⁰)-- or --CON(R¹⁰)CO-- (wherein R⁹ is a hydrogen atom, a lower alkyl group, a formimidoyl group or an acetoimidoyl group, and R¹⁰ is a hydrogen atom or a lower alkyl group), and each of m and n which may be the same or different, is an integer of from 0 to 3, provided that m and n are not simultaneously 0, a compound of the formula: ##STR11## wherein R is a hydrogen atom or a methyl group, A is ═NR⁹, --CON(R¹⁰)-- or --CON(R¹⁰)CO-- (wherein R⁹ is a hydrogen atom, a lower alkyl group, a formimidoyl group or an acetoimidoyl group, R¹⁰ is a hydrogen atom or a lower alkyl group), and each of m, n and p which may be the same or different, is an integer of from 0 to 3, provided that m and n are not simultaneously 0, and a compound of the formula: ##STR12## wherein R is a hydrogen atom or a methyl group, R¹ is a hydrogen atom or a negative charge, A² is ═NR⁹ ═N⁺ (R¹¹)R¹², --CON(R¹⁰)-- or --CON(R¹⁰)CO-- (wherein R⁹ is a hydrogen atom, a lower alkyl group, a formimidoyl group or an acetoimidoyl group, R¹⁰ is a hydrogen atom or a lower alkyl group, and each of R¹¹ and R¹² which may be the same or different, is a lower alkyl group), and each of m, n and p which may be the same or different, is an integer of from 0 to 3, provided that m and n are not simultaneously 0.

Among the compounds of the formulas (I-a) and (I-b), those wherein A¹ is --CON(R¹⁰)-- or ═NR⁹, are preferred

Among the compounds of the formula (I-c), those therein A² is ═NR⁹, ═N⁺ (R¹¹)R¹² or --CON(R¹⁰)-, are preferred.

Specific examples of the compound of the formula (I) include, for example, the following compounds.

                  TABLE 1                                                          ______________________________________                                          ##STR13##                                                                      No.        R.sup.1                                                                              ##STR14##                                                    ______________________________________                                          1         H                                                                                     ##STR15##                                                     2         H                                                                                     ##STR16##                                                     3         H                                                                                     ##STR17##                                                     4         H                                                                                     ##STR18##                                                     5         H                                                                                     ##STR19##                                                     6         H                                                                                     ##STR20##                                                     7         H                                                                                     ##STR21##                                                     8         H                                                                                     ##STR22##                                                     9         H                                                                                     ##STR23##                                                    10         H                                                                                     ##STR24##                                                    11         H                                                                                     ##STR25##                                                    12         H                                                                                     ##STR26##                                                    13         H                                                                                     ##STR27##                                                    14         H                                                                                     ##STR28##                                                    15         H                                                                                     ##STR29##                                                    16         H                                                                                     ##STR30##                                                    17         H                                                                                     ##STR31##                                                    18         H                                                                                     ##STR32##                                                    19         H                                                                                     ##STR33##                                                    20         H                                                                                     ##STR34##                                                    21         H                                                                                     ##STR35##                                                    22         H                                                                                     ##STR36##                                                    23         H                                                                                     ##STR37##                                                    24         H                                                                                     ##STR38##                                                    ______________________________________                                    

                  TABLE 2                                                          ______________________________________                                          No.        R.sup.1                                                                              ##STR39##                                                    ______________________________________                                         25         H                                                                                     ##STR40##                                                    26         H                                                                                     ##STR41##                                                    27         H                                                                                     ##STR42##                                                    28         H                                                                                     ##STR43##                                                    29         H                                                                                     ##STR44##                                                    30         H                                                                                     ##STR45##                                                    31         H                                                                                     ##STR46##                                                    32         H                                                                                     ##STR47##                                                    33         H                                                                                     ##STR48##                                                    34         H                                                                                     ##STR49##                                                    35         H                                                                                     ##STR50##                                                    36         H                                                                                     ##STR51##                                                    37         H                                                                                     ##STR52##                                                    38         H                                                                                     ##STR53##                                                    39         H                                                                                     ##STR54##                                                    40         H                                                                                     ##STR55##                                                    41         H                                                                                     ##STR56##                                                    42         H                                                                                     ##STR57##                                                    43         H                                                                                     ##STR58##                                                    44         H                                                                                     ##STR59##                                                    45         H                                                                                     ##STR60##                                                    46         H                                                                                     ##STR61##                                                    47         H                                                                                     ##STR62##                                                    48         H                                                                                     ##STR63##                                                    49         H                                                                                     ##STR64##                                                    50         H                                                                                     ##STR65##                                                    51         H                                                                                     ##STR66##                                                    52         H                                                                                     ##STR67##                                                    ______________________________________                                    

                  TABLE 3                                                          ______________________________________                                          No.      R.sup.1                                                                                 ##STR68##                                                   ______________________________________                                         53       H                                                                                        ##STR69##                                                   54       H                                                                                        ##STR70##                                                   55       H                                                                                        ##STR71##                                                   56       H                                                                                        ##STR72##                                                   57       H                                                                                        ##STR73##                                                   58       H                                                                                        ##STR74##                                                   59       H                                                                                        ##STR75##                                                   60       H                                                                                        ##STR76##                                                   61       H                                                                                        ##STR77##                                                   62       H                                                                                        ##STR78##                                                   63       Negative charge                                                                          ##STR79##                                                   64       Negative charge                                                                          ##STR80##                                                   65       Negative charge                                                                          ##STR81##                                                   66       Negative charge                                                                          ##STR82##                                                   67       Negative charge                                                                          ##STR83##                                                   68       Negative charge                                                                          ##STR84##                                                   69       H                                                                                        ##STR85##                                                   70       H                                                                                        ##STR86##                                                   71       H                                                                                        ##STR87##                                                   72       H                                                                                        ##STR88##                                                   73       H                                                                                        ##STR89##                                                   74       Negative charge                                                                          ##STR90##                                                   75       Negative charge                                                                          ##STR91##                                                   76       H                                                                                        ##STR92##                                                   77       H                                                                                        ##STR93##                                                   78       Negative charge                                                                          ##STR94##                                                   ______________________________________                                    

                  TABLE 4                                                          ______________________________________                                          No.         R.sup.1                                                                              ##STR95##                                                   ______________________________________                                         79          H                                                                                     ##STR96##                                                   80          H                                                                                     ##STR97##                                                   81          H                                                                                     ##STR98##                                                   82          H                                                                                     ##STR99##                                                   83          H                                                                                     ##STR100##                                                  84          H                                                                                     ##STR101##                                                  85          H                                                                                     ##STR102##                                                  86          H                                                                                     ##STR103##                                                  87          H                                                                                     ##STR104##                                                  88          H                                                                                     ##STR105##                                                  89          H                                                                                     ##STR106##                                                  90          H                                                                                     ##STR107##                                                  91          H                                                                                     ##STR108##                                                  92          H                                                                                     ##STR109##                                                  93          H                                                                                     ##STR110##                                                  94          H                                                                                     ##STR111##                                                  95          H                                                                                     ##STR112##                                                  96          H                                                                                     ##STR113##                                                  97          H                                                                                     ##STR114##                                                  98          H                                                                                     ##STR115##                                                  99          H                                                                                     ##STR116##                                                  100         H                                                                                     ##STR117##                                                  ______________________________________                                    

                  TABLE 5                                                          ______________________________________                                          No.       R.sup.1                                                                                 ##STR118##                                                 ______________________________________                                         101       H                                                                                        ##STR119##                                                 102       H                                                                                        ##STR120##                                                 103       H                                                                                        ##STR121##                                                 104       H                                                                                        ##STR122##                                                 105       H                                                                                        ##STR123##                                                 106       H                                                                                        ##STR124##                                                 107       H                                                                                        ##STR125##                                                 108       H                                                                                        ##STR126##                                                 109       H                                                                                        ##STR127##                                                 110       H                                                                                        ##STR128##                                                 111       H                                                                                        ##STR129##                                                 112       H                                                                                        ##STR130##                                                 113       H                                                                                        ##STR131##                                                 114       Negative charge                                                                          ##STR132##                                                 115       H                                                                                        ##STR133##                                                 116       H                                                                                        ##STR134##                                                 117       H                                                                                        ##STR135##                                                 118       H                                                                                        ##STR136##                                                 119       Negative charge                                                                          ##STR137##                                                 120       Negative charge                                                                          ##STR138##                                                 121       H                                                                                        ##STR139##                                                 122       H                                                                                        ##STR140##                                                 ______________________________________                                    

                  TABLE 6                                                          ______________________________________                                          No.       R.sup.1                                                                                 ##STR141##                                                 ______________________________________                                         123       H                                                                                        ##STR142##                                                 124       H                                                                                        ##STR143##                                                 125       H                                                                                        ##STR144##                                                 126       H                                                                                        ##STR145##                                                 127       Negative charge                                                                          ##STR146##                                                 128       Negative charge                                                                          ##STR147##                                                 129       Negative charge                                                                          ##STR148##                                                 ______________________________________                                    

                  TABLE 7                                                          ______________________________________                                          ##STR149##                                                                     No.         R.sup.1                                                                              ##STR150##                                                  ______________________________________                                         130         H                                                                                     ##STR151##                                                  131         H                                                                                     ##STR152##                                                  132         H                                                                                     ##STR153##                                                  133         H                                                                                     ##STR154##                                                  134         H                                                                                     ##STR155##                                                  135         H                                                                                     ##STR156##                                                  136         H                                                                                     ##STR157##                                                  137         H                                                                                     ##STR158##                                                  138         H                                                                                     ##STR159##                                                  139         H                                                                                     ##STR160##                                                  140         H                                                                                     ##STR161##                                                  141         H                                                                                     ##STR162##                                                  142         H                                                                                     ##STR163##                                                  143         H                                                                                     ##STR164##                                                  144         H                                                                                     ##STR165##                                                  145         H                                                                                     ##STR166##                                                  146         H                                                                                     ##STR167##                                                  147         H                                                                                     ##STR168##                                                  148         H                                                                                     ##STR169##                                                  149         H                                                                                     ##STR170##                                                  150         H                                                                                     ##STR171##                                                  151         H                                                                                     ##STR172##                                                  152         H                                                                                     ##STR173##                                                  153         H                                                                                     ##STR174##                                                  ______________________________________                                    

                  TABLE 8                                                          ______________________________________                                          No.        R.sup.1                                                                               ##STR175##                                                  ______________________________________                                         154        H                                                                                      ##STR176##                                                  155        H                                                                                      ##STR177##                                                  156        H                                                                                      ##STR178##                                                  157        H                                                                                      ##STR179##                                                  158        H                                                                                      ##STR180##                                                  159        H                                                                                      ##STR181##                                                  160        H                                                                                      ##STR182##                                                  161        H                                                                                      ##STR183##                                                  162        H                                                                                      ##STR184##                                                  163        H                                                                                      ##STR185##                                                  164        H                                                                                      ##STR186##                                                  165        H                                                                                      ##STR187##                                                  166        H                                                                                      ##STR188##                                                  167        H                                                                                      ##STR189##                                                  168        H                                                                                      ##STR190##                                                  169        H                                                                                      ##STR191##                                                  170        H                                                                                      ##STR192##                                                  171        H                                                                                      ##STR193##                                                  172        H                                                                                      ##STR194##                                                  173        H                                                                                      ##STR195##                                                  174        H                                                                                      ##STR196##                                                  175        H                                                                                      ##STR197##                                                  176        H                                                                                      ##STR198##                                                  177        H                                                                                      ##STR199##                                                  178        H                                                                                      ##STR200##                                                  179        H                                                                                      ##STR201##                                                  180        H                                                                                      ##STR202##                                                  181        H                                                                                      ##STR203##                                                  ______________________________________                                    

                  TABLE 9                                                          ______________________________________                                          No.     R.sup.1                                                                                     ##STR204##                                               ______________________________________                                         182     H                                                                                            ##STR205##                                               183     H                                                                                            ##STR206##                                               184     H                                                                                            ##STR207##                                               185     H                                                                                            ##STR208##                                               186     H                                                                                            ##STR209##                                               187     H                                                                                            ##STR210##                                               188     H                                                                                            ##STR211##                                               189     H                                                                                            ##STR212##                                               190     H                                                                                            ##STR213##                                               191     H                                                                                            ##STR214##                                               192     Negative charge                                                                              ##STR215##                                               193     Negative charge                                                                              ##STR216##                                               194     Negative charge                                                                              ##STR217##                                               195     Negative charge                                                                              ##STR218##                                               196     Negative charge                                                                              ##STR219##                                               197     Negative charge                                                                              ##STR220##                                               198     H                                                                                            ##STR221##                                               199     H                                                                                            ##STR222##                                               200     H                                                                                            ##STR223##                                               201     H                                                                                            ##STR224##                                               202     H                                                                                            ##STR225##                                               203     Negative charge                                                                              ##STR226##                                               204     Negative charge                                                                              ##STR227##                                               205     H                                                                                            ##STR228##                                               206     H                                                                                            ##STR229##                                               207     Negative charge                                                                              ##STR230##                                               ______________________________________                                    

                  TABLE 10                                                         ______________________________________                                          No.        R.sup.1                                                                               ##STR231##                                                  ______________________________________                                         208        H                                                                                      ##STR232##                                                  209        H                                                                                      ##STR233##                                                  210        H                                                                                      ##STR234##                                                  211        H                                                                                      ##STR235##                                                  212        H                                                                                      ##STR236##                                                  213        H                                                                                      ##STR237##                                                  214        H                                                                                      ##STR238##                                                  215        H                                                                                      ##STR239##                                                  216        H                                                                                      ##STR240##                                                  217        H                                                                                      ##STR241##                                                  218        H                                                                                      ##STR242##                                                  219        H                                                                                      ##STR243##                                                  220        H                                                                                      ##STR244##                                                  221        H                                                                                      ##STR245##                                                  222        H                                                                                      ##STR246##                                                  223        H                                                                                      ##STR247##                                                  224        H                                                                                      ##STR248##                                                  225        H                                                                                      ##STR249##                                                  226        H                                                                                      ##STR250##                                                  227        H                                                                                      ##STR251##                                                  228        H                                                                                      ##STR252##                                                  229        H                                                                                      ##STR253##                                                  ______________________________________                                    

                                      TABLE 11                                     __________________________________________________________________________      No.                                                                                R.sup.1                                                                           ##STR254##    No.                                                                                R.sup.1                                                                            ##STR255##                                       __________________________________________________________________________     230 H                                                                                  ##STR256##   241 H                                                                                   ##STR257##                                       231 H                                                                                  ##STR258##   242 H                                                                                   ##STR259##                                       232 H                                                                                  ##STR260##   243 Nega- tive charge                                                                   ##STR261##                                       233 H                                                                                  ##STR262##   244 H                                                                                   ##STR263##                                       234 H                                                                                  ##STR264##   245 H                                                                                   ##STR265##                                       235 H                                                                                  ##STR266##   246 H                                                                                   ##STR267##                                       236 H                                                                                  ##STR268##   247 H                                                                                   ##STR269##                                       237 H                                                                                  ##STR270##   248 Nega- tive charge                                                                   ##STR271##                                       238 H                                                                                  ##STR272##   249 Nega- tive charge                                                                   ##STR273##                                       239 H                                                                                  ##STR274##   250 H                                                                                   ##STR275##                                       240 H                                                                                  ##STR276##   251 H                                                                                   ##STR277##                                       __________________________________________________________________________

                                      TABLE 12                                     __________________________________________________________________________      No.                                                                                R.sup.1                                                                           ##STR278##  No.                                                                                R.sup.1                                                                            ##STR279##                                         __________________________________________________________________________     252 H                                                                                  ##STR280## 256 Nega- tive charge                                                                   ##STR281##                                         253 H                                                                                  ##STR282## 257 Nega- tive charge                                                                   ##STR283##                                         254 H                                                                                  ##STR284## 258 Nega- tive charge                                                                   ##STR285##                                         255 H                                                                                  ##STR286##                                                             __________________________________________________________________________

                                      TABLE 13                                     __________________________________________________________________________      ##STR287##                                                                     No.                                                                                R.sup.1                                                                           ##STR288##   No.                                                                                R.sup.1                                                                           ##STR289##                                         __________________________________________________________________________     259 H                                                                                  ##STR290##  271 H                                                                                  ##STR291##                                         260 H                                                                                  ##STR292##  272 H                                                                                  ##STR293##                                         261 H                                                                                  ##STR294##  273 H                                                                                  ##STR295##                                         262 H                                                                                  ##STR296##  274 H                                                                                  ##STR297##                                         263 H                                                                                  ##STR298##  275 H                                                                                  ##STR299##                                         264 H                                                                                  ##STR300##  276 H                                                                                  ##STR301##                                         265 H                                                                                  ##STR302##  277 H                                                                                  ##STR303##                                         266 H                                                                                  ##STR304##  278 H                                                                                  ##STR305##                                         267 H                                                                                  ##STR306##  279 H                                                                                  ##STR307##                                         268 H                                                                                  ##STR308##  280 H                                                                                  ##STR309##                                         269 H                                                                                  ##STR310##  281 H                                                                                  ##STR311##                                         270 H                                                                                  ##STR312##  282 H                                                                                  ##STR313##                                         __________________________________________________________________________

                  TABLE 14                                                         ______________________________________                                          No.  R.sup.1                                                                              ##STR314##      No.  R.sup.1                                                                            ##STR315##                                ______________________________________                                         283  H                                                                                     ##STR316##     297  H                                                                                   ##STR317##                                284  H                                                                                     ##STR318##     298  H                                                                                   ##STR319##                                285  H                                                                                     ##STR320##     299  H                                                                                   ##STR321##                                286  H                                                                                     ##STR322##     300  H                                                                                   ##STR323##                                287  H                                                                                     ##STR324##     301  H                                                                                   ##STR325##                                288  H                                                                                     ##STR326##     302  H                                                                                   ##STR327##                                289  H                                                                                     ##STR328##     303  H                                                                                   ##STR329##                                290  H                                                                                     ##STR330##     304  H                                                                                   ##STR331##                                291  H                                                                                     ##STR332##     305  H                                                                                   ##STR333##                                292  H                                                                                     ##STR334##     306  H                                                                                   ##STR335##                                293  H                                                                                     ##STR336##     307  H                                                                                   ##STR337##                                294  H                                                                                     ##STR338##     308  H                                                                                   ##STR339##                                295  H                                                                                     ##STR340##     309  H                                                                                   ##STR341##                                296  H                                                                                     ##STR342##     310  H                                                                                   ##STR343##                                ______________________________________                                    

                                      TABLE 15                                     __________________________________________________________________________      No.                                                                               R.sup.1                                                                            ##STR344##    No.                                                                               R.sup.1                                                                            ##STR345##                                        __________________________________________________________________________     311                                                                               H                                                                                   ##STR346##   324                                                                               Nega- tive charge                                                                   ##STR347##                                        312                                                                               H                                                                                   ##STR348##   325                                                                               Nega- tive charge                                                                   ##STR349##                                        313                                                                               H                                                                                   ##STR350##   326                                                                               Nega- tive charge                                                                   ##STR351##                                        314                                                                               H                                                                                   ##STR352##   327                                                                               H                                                                                   ##STR353##                                        315                                                                               H                                                                                   ##STR354##   328                                                                               H                                                                                   ##STR355##                                        316                                                                               H                                                                                   ##STR356##   329                                                                               H                                                                                   ##STR357##                                        317                                                                               H                                                                                   ##STR358##   330                                                                               H                                                                                   ##STR359##                                        318                                                                               H                                                                                   ##STR360##   331                                                                               H                                                                                   ##STR361##                                        319                                                                               H                                                                                   ##STR362##   332                                                                               Nega- tive charge                                                                   ##STR363##                                        320                                                                               H                                                                                   ##STR364##   333                                                                               Nega- tive charge                                                                   ##STR365##                                        321                                                                               Nega- tive charge                                                                   ##STR366##   334                                                                               H                                                                                   ##STR367##                                        322                                                                               Nega- tive charge                                                                   ##STR368##   335                                                                               H                                                                                   ##STR369##                                        323                                                                               Nega- tive charge                                                                   ##STR370##   336                                                                               Nega- tive charge                                                                   ##STR371##                                        __________________________________________________________________________

                                      TABLE 16                                     __________________________________________________________________________      No.                                                                                R.sup.1                                                                           ##STR372##  No.                                                                                R.sup.1                                                                           ##STR373##                                          __________________________________________________________________________     337 H                                                                                  ##STR374## 348 H                                                                                  ##STR375##                                          338 H                                                                                  ##STR376## 349 H                                                                                  ##STR377##                                          339 H                                                                                  ##STR378## 350 H                                                                                  ##STR379##                                          340 H                                                                                  ##STR380## 351 H                                                                                  ##STR381##                                          341 H                                                                                  ##STR382## 352 H                                                                                  ##STR383##                                          342 H                                                                                  ##STR384## 353 H                                                                                  ##STR385##                                          343 H                                                                                  ##STR386## 354 H                                                                                  ##STR387##                                          344 H                                                                                  ##STR388## 355 H                                                                                  ##STR389##                                          345 H                                                                                  ##STR390## 356 H                                                                                  ##STR391##                                          346 H                                                                                  ##STR392## 357 H                                                                                  ##STR393##                                          347 H                                                                                  ##STR394## 358 H                                                                                  ##STR395##                                          __________________________________________________________________________

                                      TABLE 17                                     __________________________________________________________________________      No.                                                                                R.sup.1                                                                           ##STR396##    No.                                                                                R.sup.1                                                                             ##STR397##                                      __________________________________________________________________________     359 H                                                                                  ##STR398##   370 H                                                                                    ##STR399##                                      360 H                                                                                  ##STR400##   371 H                                                                                    ##STR401##                                      361 H                                                                                  ##STR402##   372 Nega- tive charge                                                                    ##STR403##                                      362 H                                                                                  ##STR404##   373 H                                                                                    ##STR405##                                      363 H                                                                                  ##STR406##   374 H                                                                                    ##STR407##                                      364 H                                                                                  ##STR408##   375 H                                                                                    ##STR409##                                      365 H                                                                                  ##STR410##   376 H                                                                                    ##STR411##                                      366 H                                                                                  ##STR412##   377 Nega- tive charge                                                                    ##STR413##                                      367 H                                                                                  ##STR414##   378 Nega- tive charge                                                                    ##STR415##                                      368 H                                                                                  ##STR416##   379 H                                                                                    ##STR417##                                      369 H                                                                                  ##STR418##   380 H                                                                                    ##STR419##                                      __________________________________________________________________________

                                      TABLE 18                                     __________________________________________________________________________      No.                                                                                R.sup.1                                                                           ##STR420##  No.                                                                                R.sup.1                                                                             ##STR421##                                        __________________________________________________________________________     381 H                                                                                  ##STR422## 385 Nega- tive charge                                                                    ##STR423##                                        382 H                                                                                  ##STR424## 386 Nega- tive charge                                                                    ##STR425##                                        383 H                                                                                  ##STR426## 387 Nega-  tive charge                                                                   ##STR427##                                        384 H                                                                                  ##STR428##                                                             __________________________________________________________________________

                                      TABLE 19                                     __________________________________________________________________________      ##STR429##                                                                     No.                                                                                R.sup.1                                                                           ##STR430##   No.                                                                                R.sup.1                                                                           ##STR431##                                         __________________________________________________________________________     388 H                                                                                  ##STR432##  400 H                                                                                  ##STR433##                                         389 H                                                                                  ##STR434##  401 H                                                                                  ##STR435##                                         390 H                                                                                  ##STR436##  402 H                                                                                  ##STR437##                                         391 H                                                                                  ##STR438##  403 H                                                                                  ##STR439##                                         392 H                                                                                  ##STR440##  404 H                                                                                  ##STR441##                                         393 H                                                                                  ##STR442##  405 H                                                                                  ##STR443##                                         394 H                                                                                  ##STR444##  406 H                                                                                  ##STR445##                                         395 H                                                                                  ##STR446##  407 H                                                                                  ##STR447##                                         396 H                                                                                  ##STR448##  408 H                                                                                  ##STR449##                                         397 H                                                                                  ##STR450##  409 H                                                                                  ##STR451##                                         398 H                                                                                  ##STR452##  410 H                                                                                  ##STR453##                                         399 H                                                                                  ##STR454##  411 H                                                                                  ##STR455##                                         __________________________________________________________________________

                  TABLE 20                                                         ______________________________________                                          No.  R.sup.1                                                                              ##STR456##      No.  R.sup.1                                                                            ##STR457##                                ______________________________________                                         412  H                                                                                     ##STR458##     426  H                                                                                   ##STR459##                                413  H                                                                                     ##STR460##     427  H                                                                                   ##STR461##                                414  H                                                                                     ##STR462##     428  H                                                                                   ##STR463##                                415  H                                                                                     ##STR464##     429  H                                                                                   ##STR465##                                416  H                                                                                     ##STR466##     430  H                                                                                   ##STR467##                                417  H                                                                                     ##STR468##     431  H                                                                                   ##STR469##                                418  H                                                                                     ##STR470##     432  H                                                                                   ##STR471##                                419  H                                                                                     ##STR472##     433  H                                                                                   ##STR473##                                420  H                                                                                     ##STR474##     434  H                                                                                   ##STR475##                                421  H                                                                                     ##STR476##     435  H                                                                                   ##STR477##                                422  H                                                                                     ##STR478##     436  H                                                                                   ##STR479##                                423  H                                                                                     ##STR480##     437  H                                                                                   ##STR481##                                424  H                                                                                     ##STR482##     438  H                                                                                   ##STR483##                                425  H                                                                                     ##STR484##     439  H                                                                                   ##STR485##                                ______________________________________                                    

                                      TABLE 21                                     __________________________________________________________________________      No.                                                                               R.sup.1                                                                            ##STR486##   No.                                                                               R.sup.1                                                                            ##STR487##                                         __________________________________________________________________________     440                                                                               H                                                                                   ##STR488##  453                                                                               Nega- tive charge                                                                   ##STR489##                                         441                                                                               H                                                                                   ##STR490##  454                                                                               Nega- tive charge                                                                   ##STR491##                                         442                                                                               H                                                                                   ##STR492##  455                                                                               Nega- tive charge                                                                   ##STR493##                                         443                                                                               H                                                                                   ##STR494##  456                                                                               H                                                                                   ##STR495##                                         444                                                                               H                                                                                   ##STR496##  457                                                                               H                                                                                   ##STR497##                                         445                                                                               H                                                                                   ##STR498##  458                                                                               H                                                                                   ##STR499##                                         446                                                                               H                                                                                   ##STR500##  459                                                                               H                                                                                   ##STR501##                                         447                                                                               H                                                                                   ##STR502##  460                                                                               H                                                                                   ##STR503##                                         448                                                                               H                                                                                   ##STR504##  461                                                                               Nega- tive charge                                                                   ##STR505##                                         449                                                                               H                                                                                   ##STR506##  462                                                                               Nega- tive charge                                                                   ##STR507##                                         450                                                                               Nega- tive charge                                                                   ##STR508##  463                                                                               H                                                                                   ##STR509##                                         451                                                                               Nega- tive charge                                                                   ##STR510##  464                                                                               H                                                                                   ##STR511##                                         452                                                                               Nega- tive charge                                                                   ##STR512##  465                                                                               Nega- tive charge                                                                   ##STR513##                                         __________________________________________________________________________

                  TABLE 22                                                         ______________________________________                                          No.        R.sup.1                                                                               ##STR514##                                                  ______________________________________                                         466        H                                                                                      ##STR515##                                                  467        H                                                                                      ##STR516##                                                  468        H                                                                                      ##STR517##                                                  469        H                                                                                      ##STR518##                                                  470        H                                                                                      ##STR519##                                                  471        H                                                                                      ##STR520##                                                  472        H                                                                                      ##STR521##                                                  473        H                                                                                      ##STR522##                                                  474        H                                                                                      ##STR523##                                                  475        H                                                                                      ##STR524##                                                  476        H                                                                                      ##STR525##                                                  477        H                                                                                      ##STR526##                                                  478        H                                                                                      ##STR527##                                                  479        H                                                                                      ##STR528##                                                  480        H                                                                                      ##STR529##                                                  481        H                                                                                      ##STR530##                                                  482        H                                                                                      ##STR531##                                                  483        H                                                                                      ##STR532##                                                  484        H                                                                                      ##STR533##                                                  485        H                                                                                      ##STR534##                                                  486        H                                                                                      ##STR535##                                                  487        H                                                                                      ##STR536##                                                  ______________________________________                                    

                  TABLE 23                                                         ______________________________________                                          No.       R.sup.1                                                                                 ##STR537##                                                 ______________________________________                                         488       H                                                                                        ##STR538##                                                 489       H                                                                                        ##STR539##                                                 490       H                                                                                        ##STR540##                                                 491       H                                                                                        ##STR541##                                                 492       H                                                                                        ##STR542##                                                 493       H                                                                                        ##STR543##                                                 494       H                                                                                        ##STR544##                                                 495       H                                                                                        ##STR545##                                                 496       H                                                                                        ##STR546##                                                 497       H                                                                                        ##STR547##                                                 498       H                                                                                        ##STR548##                                                 499       H                                                                                        ##STR549##                                                 500       H                                                                                        ##STR550##                                                 501       Negative charge                                                                          ##STR551##                                                 502       H                                                                                        ##STR552##                                                 503       H                                                                                        ##STR553##                                                 504       H                                                                                        ##STR554##                                                 505       H                                                                                        ##STR555##                                                 506       Negative charge                                                                          ##STR556##                                                 507       Negative charge                                                                          ##STR557##                                                 508       H                                                                                        ##STR558##                                                 509       H                                                                                        ##STR559##                                                 ______________________________________                                    

                  TABLE 24                                                         ______________________________________                                          No.       R.sup.1                                                                                 ##STR560##                                                 ______________________________________                                         510       H                                                                                        ##STR561##                                                 511       H                                                                                        ##STR562##                                                 512       H                                                                                        ##STR563##                                                 513       H                                                                                        ##STR564##                                                 514       Negative charge                                                                          ##STR565##                                                 515       Negative charge                                                                          ##STR566##                                                 516       Negative charge                                                                          ##STR567##                                                 ______________________________________                                    

                  TABLE 25                                                         ______________________________________                                          ##STR568##                                                                     No.       R.sup.1                                                                               ##STR569##                                                   ______________________________________                                         517       H                                                                                      ##STR570##                                                   518       H                                                                                      ##STR571##                                                   519       H                                                                                      ##STR572##                                                   520       H                                                                                      ##STR573##                                                   521       H                                                                                      ##STR574##                                                   522       H                                                                                      ##STR575##                                                   523       H                                                                                      ##STR576##                                                   524       H                                                                                      ##STR577##                                                   525       H                                                                                      ##STR578##                                                   526       H                                                                                      ##STR579##                                                   527       H                                                                                      ##STR580##                                                   528       H                                                                                      ##STR581##                                                   529       H                                                                                      ##STR582##                                                   530       H                                                                                      ##STR583##                                                   531       H                                                                                      ##STR584##                                                   532       H                                                                                      ##STR585##                                                   533       H                                                                                      ##STR586##                                                   534       H                                                                                      ##STR587##                                                   535       H                                                                                      ##STR588##                                                   536       H                                                                                      ##STR589##                                                   537       H                                                                                      ##STR590##                                                   538       H                                                                                      ##STR591##                                                   539       H                                                                                      ##STR592##                                                   540       H                                                                                      ##STR593##                                                   ______________________________________                                    

                  TABLE 26                                                         ______________________________________                                          No.        R.sup.1                                                                              ##STR594##                                                   ______________________________________                                         541        H                                                                                     ##STR595##                                                   542        H                                                                                     ##STR596##                                                   543        H                                                                                     ##STR597##                                                   544        H                                                                                     ##STR598##                                                   545        H                                                                                     ##STR599##                                                   546        H                                                                                     ##STR600##                                                   547        H                                                                                     ##STR601##                                                   548        H                                                                                     ##STR602##                                                   549        H                                                                                     ##STR603##                                                   550        H                                                                                     ##STR604##                                                   551        H                                                                                     ##STR605##                                                   552        H                                                                                     ##STR606##                                                   553        H                                                                                     ##STR607##                                                   554        H                                                                                     ##STR608##                                                   555        H                                                                                     ##STR609##                                                   556        H                                                                                     ##STR610##                                                   557        H                                                                                     ##STR611##                                                   558        H                                                                                     ##STR612##                                                   559        H                                                                                     ##STR613##                                                   560        H                                                                                     ##STR614##                                                   561        H                                                                                     ##STR615##                                                   562        H                                                                                     ##STR616##                                                   563        H                                                                                     ##STR617##                                                   564        H                                                                                     ##STR618##                                                   565        H                                                                                     ##STR619##                                                   566        H                                                                                     ##STR620##                                                   567        H                                                                                     ##STR621##                                                   568        H                                                                                     ##STR622##                                                   ______________________________________                                    

                  TABLE 27                                                         ______________________________________                                          No.       R.sup.1                                                                                 ##STR623##                                                 ______________________________________                                         569       H                                                                                        ##STR624##                                                 570       H                                                                                        ##STR625##                                                 571       H                                                                                        ##STR626##                                                 572       H                                                                                        ##STR627##                                                 573       H                                                                                        ##STR628##                                                 574       H                                                                                        ##STR629##                                                 575       H                                                                                        ##STR630##                                                 576       H                                                                                        ##STR631##                                                 577       H                                                                                        ##STR632##                                                 578       H                                                                                        ##STR633##                                                 579       Negative charge                                                                          ##STR634##                                                 580       Negative charge                                                                          ##STR635##                                                 581       Negative charge                                                                          ##STR636##                                                 582       Negative charge                                                                          ##STR637##                                                 583       Negative charge                                                                          ##STR638##                                                 584       Negative  charge                                                                         ##STR639##                                                 585       H                                                                                        ##STR640##                                                 586       H                                                                                        ##STR641##                                                 587       H                                                                                        ##STR642##                                                 588       H                                                                                        ##STR643##                                                 589       H                                                                                        ##STR644##                                                 590       Negative charge                                                                          ##STR645##                                                 591       Negative charge                                                                          ##STR646##                                                 592       H                                                                                        ##STR647##                                                 593       H                                                                                        ##STR648##                                                 594       Negative charge                                                                          ##STR649##                                                 ______________________________________                                    

                  TABLE 28                                                         ______________________________________                                          No.        R.sup.1                                                                              ##STR650##                                                   ______________________________________                                         595        H                                                                                     ##STR651##                                                   596        H                                                                                     ##STR652##                                                   597        H                                                                                     ##STR653##                                                   598        H                                                                                     ##STR654##                                                   599        H                                                                                     ##STR655##                                                   600        H                                                                                     ##STR656##                                                   601        H                                                                                     ##STR657##                                                   602        H                                                                                     ##STR658##                                                   603        H                                                                                     ##STR659##                                                   604        H                                                                                     ##STR660##                                                   605        H                                                                                     ##STR661##                                                   606        H                                                                                     ##STR662##                                                   607        H                                                                                     ##STR663##                                                   608        H                                                                                     ##STR664##                                                   609        H                                                                                     ##STR665##                                                   610        H                                                                                     ##STR666##                                                   611        H                                                                                     ##STR667##                                                   612        H                                                                                     ##STR668##                                                   613        H                                                                                     ##STR669##                                                   614        H                                                                                     ##STR670##                                                   615        H                                                                                     ##STR671##                                                   616        H                                                                                     ##STR672##                                                   ______________________________________                                    

                  TABLE 29                                                         ______________________________________                                          No.       R.sup.1                                                                                 ##STR673##                                                 ______________________________________                                         617       H                                                                                        ##STR674##                                                 618       H                                                                                        ##STR675##                                                 619       H                                                                                        ##STR676##                                                 620       H                                                                                        ##STR677##                                                 621       H                                                                                        ##STR678##                                                 622       H                                                                                        ##STR679##                                                 623       H                                                                                        ##STR680##                                                 624       H                                                                                        ##STR681##                                                 625       H                                                                                        ##STR682##                                                 626       H                                                                                        ##STR683##                                                 627       H                                                                                        ##STR684##                                                 628       H                                                                                        ##STR685##                                                 629       H                                                                                        ##STR686##                                                 630       Negative charge                                                                          ##STR687##                                                 631       H                                                                                        ##STR688##                                                 632       H                                                                                        ##STR689##                                                 633       H                                                                                        ##STR690##                                                 634       H                                                                                        ##STR691##                                                 635       Negative charge                                                                          ##STR692##                                                 636       Negative charge                                                                          ##STR693##                                                 637       H                                                                                        ##STR694##                                                 638       H                                                                                        ##STR695##                                                 ______________________________________                                    

                  TABLE 30                                                         ______________________________________                                          No.       R.sup.1                                                                                 ##STR696##                                                 ______________________________________                                         639       H                                                                                        ##STR697##                                                 640       H                                                                                        ##STR698##                                                 641       H                                                                                        ##STR699##                                                 642       H                                                                                        ##STR700##                                                 643       Negative charge                                                                          ##STR701##                                                 644       Negative charge                                                                          ##STR702##                                                 645       Negative charge                                                                          ##STR703##                                                 ______________________________________                                    

                  TABLE 31                                                         ______________________________________                                          ##STR704##                                                                     No.         R.sup.1                                                                               ##STR705##                                                 ______________________________________                                         646         H                                                                                      ##STR706##                                                 647         H                                                                                      ##STR707##                                                 648         H                                                                                      ##STR708##                                                 649         H                                                                                      ##STR709##                                                 650         H                                                                                      ##STR710##                                                 651         H                                                                                      ##STR711##                                                 652         H                                                                                      ##STR712##                                                 653         H                                                                                      ##STR713##                                                 654         H                                                                                      ##STR714##                                                 655         H                                                                                      ##STR715##                                                 656         H                                                                                      ##STR716##                                                 657         H                                                                                      ##STR717##                                                 658         H                                                                                      ##STR718##                                                 659         H                                                                                      ##STR719##                                                 660         H                                                                                      ##STR720##                                                 661         H                                                                                      ##STR721##                                                 662         H                                                                                      ##STR722##                                                 663         H                                                                                      ##STR723##                                                 664         H                                                                                      ##STR724##                                                 665         H                                                                                      ##STR725##                                                 666         H                                                                                      ##STR726##                                                 667         H                                                                                      ##STR727##                                                 668         H                                                                                      ##STR728##                                                 669         H                                                                                      ##STR729##                                                 ______________________________________                                    

                  TABLE 32                                                         ______________________________________                                          No.         R.sup.1                                                                               ##STR730##                                                 ______________________________________                                         670         H                                                                                      ##STR731##                                                 671         H                                                                                      ##STR732##                                                 672         H                                                                                      ##STR733##                                                 673         H                                                                                      ##STR734##                                                 674         H                                                                                      ##STR735##                                                 675         H                                                                                      ##STR736##                                                 676         H                                                                                      ##STR737##                                                 677         H                                                                                      ##STR738##                                                 678         H                                                                                      ##STR739##                                                 679         H                                                                                      ##STR740##                                                 680         H                                                                                      ##STR741##                                                 681         H                                                                                      ##STR742##                                                 682         H                                                                                      ##STR743##                                                 683         H                                                                                      ##STR744##                                                 684         H                                                                                      ##STR745##                                                 685         H                                                                                      ##STR746##                                                 686         H                                                                                      ##STR747##                                                 687         H                                                                                      ##STR748##                                                 688         H                                                                                      ##STR749##                                                 689         H                                                                                      ##STR750##                                                 690         H                                                                                      ##STR751##                                                 691         H                                                                                      ##STR752##                                                 692         H                                                                                      ##STR753##                                                 693         H                                                                                      ##STR754##                                                 694         H                                                                                      ##STR755##                                                 695         H                                                                                      ##STR756##                                                 696         H                                                                                      ##STR757##                                                 697         H                                                                                      ##STR758##                                                 ______________________________________                                    

                                      TABLE 33                                     __________________________________________________________________________      No.                                                                               R.sup.1                                                                             ##STR759##   No.                                                                               R.sup.1                                                                             ##STR760##                                       __________________________________________________________________________     698                                                                               H                                                                                    ##STR761##  711                                                                               Nega- tive charge                                                                    ##STR762##                                       699                                                                               H                                                                                    ##STR763##  712                                                                               Nega- tive charge                                                                    ##STR764##                                       700                                                                               H                                                                                    ##STR765##  713                                                                               Nega- tive charge                                                                    ##STR766##                                       701                                                                               H                                                                                    ##STR767##  714                                                                               H                                                                                    ##STR768##                                       702                                                                               H                                                                                    ##STR769##  715                                                                               H                                                                                    ##STR770##                                       703                                                                               H                                                                                    ##STR771##  716                                                                               H                                                                                    ##STR772##                                       704                                                                               H                                                                                    ##STR773##  717                                                                               H                                                                                    ##STR774##                                       705                                                                               H                                                                                    ##STR775##  718                                                                               H                                                                                    ##STR776##                                       706                                                                               H                                                                                    ##STR777##  719                                                                               Nega- tive charge                                                                    ##STR778##                                       707                                                                               H                                                                                    ##STR779##  720                                                                               Nega- tive charge                                                                    ##STR780##                                       708                                                                               Nega- tive charge                                                                    ##STR781##  721                                                                               H                                                                                    ##STR782##                                       709                                                                               Nega- tive charge                                                                    ##STR783##  722                                                                               H                                                                                    ##STR784##                                       710                                                                               Nega- tive charge                                                                    ##STR785##  723                                                                               Nega- tive charge                                                                    ##STR786##                                       __________________________________________________________________________

                                      TABLE 34                                     __________________________________________________________________________      No.                                                                               R.sup.1                                                                           ##STR787##    No.                                                                               R.sup.1                                                                           ##STR788##                                          __________________________________________________________________________     724                                                                               H                                                                                  ##STR789##   735                                                                               H                                                                                  ##STR790##                                          725                                                                               H                                                                                  ##STR791##   736                                                                               H                                                                                  ##STR792##                                          726                                                                               H                                                                                  ##STR793##   737                                                                               H                                                                                  ##STR794##                                          727                                                                               H                                                                                  ##STR795##   738                                                                               H                                                                                  ##STR796##                                          728                                                                               H                                                                                  ##STR797##   739                                                                               H                                                                                  ##STR798##                                          729                                                                               H                                                                                  ##STR799##   740                                                                               H                                                                                  ##STR800##                                          730                                                                               H                                                                                  ##STR801##   741                                                                               H                                                                                  ##STR802##                                          731                                                                               H                                                                                  ##STR803##   742                                                                               H                                                                                  ##STR804##                                          732                                                                               H                                                                                  ##STR805##   743                                                                               H                                                                                  ##STR806##                                          733                                                                               H                                                                                  ##STR807##   744                                                                               H                                                                                  ##STR808##                                          734                                                                               H                                                                                  ##STR809##   745                                                                               H                                                                                  ##STR810##                                          __________________________________________________________________________

                                      TABLE 35                                     __________________________________________________________________________      No.                                                                               R.sup.1                                                                          ##STR811##   No.                                                                               R.sup.1                                                                             ##STR812##                                          __________________________________________________________________________     746                                                                               H                                                                                 ##STR813##  757                                                                               H                                                                                    ##STR814##                                          747                                                                               H                                                                                 ##STR815##  758                                                                               H                                                                                    ##STR816##                                          748                                                                               H                                                                                 ##STR817##  759                                                                               Nega- tive charge                                                                    ##STR818##                                          749                                                                               H                                                                                 ##STR819##  760                                                                               H                                                                                    ##STR820##                                          750                                                                               H                                                                                 ##STR821##  761                                                                               H                                                                                    ##STR822##                                          751                                                                               H                                                                                 ##STR823##  762                                                                               H                                                                                    ##STR824##                                          752                                                                               H                                                                                 ##STR825##  763                                                                               H                                                                                    ##STR826##                                          753                                                                               H                                                                                 ##STR827##  764                                                                               Nega- tive charge                                                                    ##STR828##                                          754                                                                               H                                                                                 ##STR829##  765                                                                               Nega- tive charge                                                                    ##STR830##                                          755                                                                               H                                                                                 ##STR831##  766                                                                               H                                                                                    ##STR832##                                          756                                                                               H                                                                                 ##STR833##  767                                                                               H                                                                                    ##STR834##                                          __________________________________________________________________________

                                      TABLE 36                                     __________________________________________________________________________      No.                                                                               R.sup.1                                                                           ##STR835##  No.                                                                               R.sup.1                                                                              ##STR836##                                         __________________________________________________________________________     768                                                                               H                                                                                  ##STR837## 772                                                                               Nega- tive charge                                                                     ##STR838##                                         769                                                                               H                                                                                  ##STR839## 773                                                                               Nega- tive charge                                                                     ##STR840##                                         770                                                                               H                                                                                  ##STR841## 774                                                                               Nega- tive charge                                                                     ##STR842##                                         771                                                                               H                                                                                  ##STR843##                                                              __________________________________________________________________________

                                      TABLE 37                                     __________________________________________________________________________      ##STR844##                                                                     No.                                                                                R.sup.1                                                                           ##STR845##   No.                                                                                R.sup.1                                                                           ##STR846##                                         __________________________________________________________________________     775 H                                                                                  ##STR847##  787 H                                                                                  ##STR848##                                         776 H                                                                                  ##STR849##  788 H                                                                                  ##STR850##                                         777 H                                                                                  ##STR851##  789 H                                                                                  ##STR852##                                         778 H                                                                                  ##STR853##  790 H                                                                                  ##STR854##                                         779 H                                                                                  ##STR855##  791 H                                                                                  ##STR856##                                         780 H                                                                                  ##STR857##  792 H                                                                                  ##STR858##                                         781 H                                                                                  ##STR859##  793 H                                                                                  ##STR860##                                         782 H                                                                                  ##STR861##  794 H                                                                                  ##STR862##                                         783 H                                                                                  ##STR863##  795 H                                                                                  ##STR864##                                         784 H                                                                                  ##STR865##  796 H                                                                                  ##STR866##                                         785 H                                                                                  ##STR867##  797 H                                                                                  ##STR868##                                         786 H                                                                                  ##STR869##  798 H                                                                                  ##STR870##                                         __________________________________________________________________________

                                      TABLE 38                                     __________________________________________________________________________      No.                                                                                R.sup.1                                                                           ##STR871##   No.                                                                                R.sup.1                                                                           ##STR872##                                         __________________________________________________________________________     799 H                                                                                  ##STR873##  813 H                                                                                  ##STR874##                                         800 H                                                                                  ##STR875##  814 H                                                                                  ##STR876##                                         801 H                                                                                  ##STR877##  815 H                                                                                  ##STR878##                                         802 H                                                                                  ##STR879##  816 H                                                                                  ##STR880##                                         803 H                                                                                  ##STR881##  817 H                                                                                  ##STR882##                                         804 H                                                                                  ##STR883##  818 H                                                                                  ##STR884##                                         805 H                                                                                  ##STR885##  819 H                                                                                  ##STR886##                                         806 H                                                                                  ##STR887##  820 H                                                                                  ##STR888##                                         807 H                                                                                  ##STR889##  821 H                                                                                  ##STR890##                                         808 H                                                                                  ##STR891##  822 H                                                                                  ##STR892##                                         809 H                                                                                  ##STR893##  823 H                                                                                  ##STR894##                                         810 H                                                                                  ##STR895##  824 H                                                                                  ##STR896##                                         811 H                                                                                  ##STR897##  825 H                                                                                  ##STR898##                                         812 H                                                                                  ##STR899##  826 H                                                                                  ##STR900##                                         __________________________________________________________________________

                                      TABLE 39                                     __________________________________________________________________________      No.                                                                                R.sup.1                                                                           ##STR901##   No.                                                                                R.sup.1                                                                           ##STR902##                                         __________________________________________________________________________     827 H                                                                                  ##STR903##  840 Nega- tive charge                                                                  ##STR904##                                         828 H                                                                                  ##STR905##  841 Nega- tive charge                                                                  ##STR906##                                         829 H                                                                                  ##STR907##  842 Nega- tive charge                                                                  ##STR908##                                         830 H                                                                                  ##STR909##  843 H                                                                                  ##STR910##                                         831 H                                                                                  ##STR911##  844 H                                                                                  ##STR912##                                         832 H                                                                                  ##STR913##  845 H                                                                                  ##STR914##                                         833 H                                                                                  ##STR915##  846 H                                                                                  ##STR916##                                         834 H                                                                                  ##STR917##  847 H                                                                                  ##STR918##                                         835 H                                                                                  ##STR919##  848 Nega- tive charge                                                                  ##STR920##                                         836 H                                                                                  ##STR921##  849 Nega- tive charge                                                                  ##STR922##                                         837 Nega- tive charge                                                                  ##STR923##  850 H                                                                                  ##STR924##                                         838 Nega- tive charge                                                                  ##STR925##  851 H                                                                                  ##STR926##                                         839 Nega- tive charge                                                                  ##STR927##  852 Nega- tive charge                                                                  ##STR928##                                         __________________________________________________________________________

                                      TABLE 40                                     __________________________________________________________________________      No.                                                                                R.sup.1                                                                           ##STR929##  No.                                                                               R.sup.1                                                                           ##STR930##                                           __________________________________________________________________________     853 H                                                                                  ##STR931## 864                                                                               H                                                                                  ##STR932##                                           854 H                                                                                  ##STR933## 865                                                                               H                                                                                  ##STR934##                                           855 H                                                                                  ##STR935## 866                                                                               H                                                                                  ##STR936##                                           856 H                                                                                  ##STR937## 867                                                                               H                                                                                  ##STR938##                                           857 H                                                                                  ##STR939## 868                                                                               H                                                                                  ##STR940##                                           858 H                                                                                  ##STR941## 869                                                                               H                                                                                  ##STR942##                                           859 H                                                                                  ##STR943## 870                                                                               H                                                                                  ##STR944##                                           860 H                                                                                  ##STR945## 871                                                                               H                                                                                  ##STR946##                                           861 H                                                                                  ##STR947## 872                                                                               H                                                                                  ##STR948##                                           862 H                                                                                  ##STR949## 873                                                                               H                                                                                  ##STR950##                                           863 H                                                                                  ##STR951## 874                                                                               H                                                                                  ##STR952##                                           __________________________________________________________________________

                                      TABLE 41                                     __________________________________________________________________________      No.                                                                               R.sup.1                                                                          ##STR953##    No.                                                                               R.sup.1                                                                            ##STR954##                                          __________________________________________________________________________     875                                                                               H                                                                                 ##STR955##   886                                                                               H                                                                                   ##STR956##                                          876                                                                               H                                                                                 ##STR957##   887                                                                               H                                                                                   ##STR958##                                          877                                                                               H                                                                                 ##STR959##   888                                                                               Nega- tive charge                                                                   ##STR960##                                          878                                                                               H                                                                                 ##STR961##   889                                                                               H                                                                                   ##STR962##                                          879                                                                               H                                                                                 ##STR963##   890                                                                               H                                                                                   ##STR964##                                          880                                                                               H                                                                                 ##STR965##   891                                                                               H                                                                                   ##STR966##                                          881                                                                               H                                                                                 ##STR967##   892                                                                               H                                                                                   ##STR968##                                          882                                                                               H                                                                                 ##STR969##   893                                                                               Nega- tive charge                                                                   ##STR970##                                          883                                                                               H                                                                                 ##STR971##   894                                                                               Nega- tive charge                                                                   ##STR972##                                          884                                                                               H                                                                                 ##STR973##   895                                                                               H                                                                                   ##STR974##                                          885                                                                               H                                                                                 ##STR975##   896                                                                               H                                                                                   ##STR976##                                          __________________________________________________________________________

                                      TABLE 42                                     __________________________________________________________________________      No.                                                                                R.sup.1                                                                          ##STR977##  No.                                                                                R.sup.1                                                                            ##STR978##                                          __________________________________________________________________________     897 H                                                                                 ##STR979## 901 Nega- tive charge                                                                   ##STR980##                                          898 H                                                                                 ##STR981## 902 Nega- tive charge                                                                   ##STR982##                                          899 H                                                                                 ##STR983## 903 Nega- tive charge                                                                   ##STR984##                                          900 H                                                                                 ##STR985##                                                              __________________________________________________________________________

                                      TABLE 43                                     __________________________________________________________________________      ##STR986##                                                                     No.                                                                                R.sup.1                                                                           ##STR987##   No.                                                                               R.sup.1                                                                           ##STR988##                                          __________________________________________________________________________     904 H                                                                                  ##STR989##  916                                                                               H                                                                                  ##STR990##                                          905 H                                                                                  ##STR991##  917                                                                               H                                                                                  ##STR992##                                          906 H                                                                                  ##STR993##  918                                                                               H                                                                                  ##STR994##                                          907 H                                                                                  ##STR995##  919                                                                               H                                                                                  ##STR996##                                          908 H                                                                                  ##STR997##  920                                                                               H                                                                                  ##STR998##                                          909 H                                                                                  ##STR999##  921                                                                               H                                                                                  ##STR1000##                                         910 H                                                                                  ##STR1001## 922                                                                               H                                                                                  ##STR1002##                                         911 H                                                                                  ##STR1003## 923                                                                               H                                                                                  ##STR1004##                                         912 H                                                                                  ##STR1005## 924                                                                               H                                                                                  ##STR1006##                                         913 H                                                                                  ##STR1007## 925                                                                               H                                                                                  ##STR1008##                                         914 H                                                                                  ##STR1009## 926                                                                               H                                                                                  ##STR1010##                                         915 H                                                                                  ##STR1011## 927                                                                               H                                                                                  ##STR1012##                                         __________________________________________________________________________

                  TABLE 44                                                         ______________________________________                                          No.      R.sup.1                                                                                     ##STR1013##                                             ______________________________________                                         928      H                                                                                            ##STR1014##                                             929      H                                                                                            ##STR1015##                                             930      H                                                                                            ##STR1016##                                             931      H                                                                                            ##STR1017##                                             932      H                                                                                            ##STR1018##                                             933      H                                                                                            ##STR1019##                                             934      H                                                                                            ##STR1020##                                             935      H                                                                                            ##STR1021##                                             936      H                                                                                            ##STR1022##                                             937      H                                                                                            ##STR1023##                                             938      H                                                                                            ##STR1024##                                             939      H                                                                                            ##STR1025##                                             940      H                                                                                            ##STR1026##                                             941      H                                                                                            ##STR1027##                                             942      H                                                                                            ##STR1028##                                             943      H                                                                                            ##STR1029##                                             944      H                                                                                            ##STR1030##                                             945      H                                                                                            ##STR1031##                                             946      H                                                                                            ##STR1032##                                             947      H                                                                                            ##STR1033##                                             948      H                                                                                            ##STR1034##                                             949      H                                                                                            ##STR1035##                                             950      H                                                                                            ##STR1036##                                             951      H                                                                                            ##STR1037##                                             952      H                                                                                            ##STR1038##                                             953      H                                                                                            ##STR1039##                                             954      H                                                                                            ##STR1040##                                             955      H                                                                                            ##STR1041##                                             ______________________________________                                    

                  TABLE 45                                                         ______________________________________                                          No.     R.sup.1                                                                                      ##STR1042##                                             ______________________________________                                         956     H                                                                                             ##STR1043##                                             957     H                                                                                             ##STR1044##                                             958     H                                                                                             ##STR1045##                                             959     H                                                                                             ##STR1046##                                             960     H                                                                                             ##STR1047##                                             961     H                                                                                             ##STR1048##                                             962     H                                                                                             ##STR1049##                                             963     H                                                                                             ##STR1050##                                             964     H                                                                                             ##STR1051##                                             965     H                                                                                             ##STR1052##                                             966     Negative charge                                                                               ##STR1053##                                             967     Negative charge                                                                               ##STR1054##                                             968     Negative charge                                                                               ##STR1055##                                             969     Negative charge                                                                               ##STR1056##                                             970     Negative charge                                                                               ##STR1057##                                             971     Negative charge                                                                               ##STR1058##                                             972     H                                                                                             ##STR1059##                                             973     H                                                                                             ##STR1060##                                             974     H                                                                                             ##STR1061##                                             975     H                                                                                             ##STR1062##                                             976     H                                                                                             ##STR1063##                                             977     Negative charge                                                                               ##STR1064##                                             978     Negative charge                                                                               ##STR1065##                                             979     H                                                                                             ##STR1066##                                             980     H                                                                                             ##STR1067##                                             981     Negative charge                                                                               ##STR1068##                                             ______________________________________                                    

                  TABLE 46                                                         ______________________________________                                          No.      R.sup.1                                                                                     ##STR1069##                                             ______________________________________                                         982      H                                                                                            ##STR1070##                                             983      H                                                                                            ##STR1071##                                             984      H                                                                                            ##STR1072##                                             985      H                                                                                            ##STR1073##                                             986      H                                                                                            ##STR1074##                                             987      H                                                                                            ##STR1075##                                             988      H                                                                                            ##STR1076##                                             989      H                                                                                            ##STR1077##                                             990      H                                                                                            ##STR1078##                                             991      H                                                                                            ##STR1079##                                             992      H                                                                                            ##STR1080##                                             998      H                                                                                            ##STR1081##                                             994      H                                                                                            ##STR1082##                                             995      H                                                                                            ##STR1083##                                             996      H                                                                                            ##STR1084##                                             997      H                                                                                            ##STR1085##                                             998      H                                                                                            ##STR1086##                                             999      H                                                                                            ##STR1087##                                             1000     H                                                                                            ##STR1088##                                             1001     H                                                                                            ##STR1089##                                             1002     H                                                                                            ##STR1090##                                             1003     H                                                                                            ##STR1091##                                             ______________________________________                                    

                  TABLE 47                                                         ______________________________________                                          No.     R.sup.1                                                                                      ##STR1092##                                             ______________________________________                                         1004    H                                                                                             ##STR1093##                                             1005    H                                                                                             ##STR1094##                                             1006    H                                                                                             ##STR1095##                                             1007    H                                                                                             ##STR1096##                                             1008    H                                                                                             ##STR1097##                                             1009    H                                                                                             ##STR1098##                                             1010    H                                                                                             ##STR1099##                                             1011    H                                                                                             ##STR1100##                                             1012    H                                                                                             ##STR1101##                                             1013    H                                                                                             ##STR1102##                                             1014    H                                                                                             ##STR1103##                                             1015    H                                                                                             ##STR1104##                                             1016    H                                                                                             ##STR1105##                                             1017    Negative charge                                                                               ##STR1106##                                             1018    H                                                                                             ##STR1107##                                             1019    H                                                                                             ##STR1108##                                             1020    H                                                                                             ##STR1109##                                             1021    H                                                                                             ##STR1110##                                             1022    Negative charge                                                                               ##STR1111##                                             1023    Negative charge                                                                               ##STR1112##                                             1024    H                                                                                             ##STR1113##                                             1025    H                                                                                             ##STR1114##                                             ______________________________________                                    

                  TABLE 48                                                         ______________________________________                                          No.     R.sup.1                                                                                      ##STR1115##                                             ______________________________________                                         1026    H                                                                                             ##STR1116##                                             1027    H                                                                                             ##STR1117##                                             1028    H                                                                                             ##STR1118##                                             1029    H                                                                                             ##STR1119##                                             1030    Negative charge                                                                               ##STR1120##                                             1031    Negative charge                                                                               ##STR1121##                                             1032    Negative charge                                                                               ##STR1122##                                             ______________________________________                                    

Among the above compounds, preferred are compounds identified by compound Nos. 5, 7, 9, 13, 14, 20, 23, 24, 26, 27, 28, 32, 34, 44, 45, 54, 63, 79, 85, 89, 90, 95, 96, 99, 100, 114, 115, 117, 119, 121, 122, 124, 125, 127, 128, 130, 132, 134, 136, 138, 140, 142, 143, 144, 149 152, 153, 155, 156, 157, 159, 161, 163, 167, 171, 173, 174, 180, 183, 192, 201, 205, 208, 214, 218, 224, 225, 227, 228, 229, 233, 237, 239, 240, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 263, 265, 267, 271, 273, 278, 281, 282, 284, 286, 290, 292, 302, 312, 313, 314, 343, 353, 354, 357, 372, 373, 375, 377, 379, 380, 382, 383, 385, 386, 388, 389, 390, 392, 393, 394, 396, 398, 400, 401, 402, 407, 410, 411, 413, 415, 417, 419, 421, 431, 438, 441, 450, 459, 463, 466, 472, 476, 482, 483, 485, 486, 495, 497, 498, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515 and 516.

Among them, particularly preferred are as follows:

34 (5R,6S)-6-[(R)-1-hydroxyethyl]-2-[(2S,4S)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid, 134 (1R,5S,6S)-2-[(2S,4S)-2-(azetidin-3-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-2-methyl-1-carbapen-2-em-3-carboxylic acid,

136 (1R,5S,6S)-2-[(2S,4S)-2-(N-formimidoylazetidin-3-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

138 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(pyrrolidin-2-yl)pyrrolidin-4-ylthio]-1- carbapen-2-em-3-carboxylic acid,

142 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(pyrrolidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

143 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methylpyrrolidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

144 (1R,5S,6S)-2-[(2S,4S)-2-(N-formimidoylpyrrolidin-3-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

149 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(piperidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

152 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(piperidin-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

153 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methylpiperidin-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

155 (1R,5S,6S)-2-[(2S,4S)-2-(2-azetidinon-4-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

157 (1R,5S,6S)-2-[(2S,4S)-2-(2-azetidinon-3-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

159 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(2-pyrrolidon-5-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

161 (1R,tS6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(2-pyrrolidon-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

163 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthiol-1-carbapen-2-em-3-carboxylic acid,

183 (1R,8S6S)-2-[(2S,4S)-2-(2-carbamoylpyrrolidin-4-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

192 (1R,5S,6S)-2-[(2S,4S)-2-(N,N-dimethyl-3-pyrrolidinio)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

208 (1R,8S6S)-2-[(2S,4S)-2-(3-amino-2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

214 (1R,8S6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(2-piperazinyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

218 (1R,8S6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(3-oxopiperazin-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

224 (1R,8S6S)-2-[(2S,4S)-2-(hexahydro-1H-1,4-diazepin-6-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

225 (1R,5S,6S)-2-[(2S,4S)-2-(hexahydro-2-oxo-1H-1,4-diazepin-6-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

240 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-2-[(2S,4S)-2N-(2-hydroxyethyl)pyrrolidin-3-yl]pyrrolidin-4-ylthio]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

243 (1R,5S,6S)-2-[(2S,4S)-2-(N,N-dimethyl-4-piperidinio)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate, 244 (1R,5S,6S)-2-[(2S,4S)-2-(hexahydroazepin-4-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

246 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methylhexahydroazepin-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid, 248 (1R,5S,6S)-2-[(2S,4S)-2-(N,N-dimethylhexahydro-4-azepinio)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate,

250 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-(2S,4S)-2-(octahydroazocin-5-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

251 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(octahydroazocin-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

253 (1R,5S6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methyloctahydroazocin-5-yl)pyrrolidin-4-ylthio]-1-carbapan-2-em-3-carboxylic acid,

254 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-(2S,4S)-2-(N-methyloctahydroazocin-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

256 (1R,5S,6S)-2-[(2S,4S)-2-(N,N-dimethyloctahydro-5-azocinio)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate,

257 (1R,5S,6S)-2-[(2S,4S)-2-(N,N-dimethyloctahydro-4-azocinio)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate,

290 (5R,6S)-6-[(R)-1-hydroxyethyl]-2-[(2R,4S)-2-(2-pyrrolidon-3-ylmethyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

392 (1R,5S,6S)-2-[(2R,4S)-2-(azetidin-3-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

394 (1R,5S,6S)-2-[(2R,4S)-2-(N-formimidoylazetidin-3-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

400 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2R,4S)-2-(pyrrolidin-3-ylmethyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

401 (1R 5S 6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2R,4S)-2-(N-methylpyrrolidin-3-ylmethyl)pyrrolidin-4ylthio]-1-carbapen-2-em-3-carboxylic acid,

402 (1R,5S,6S)-2-[(2R,4.S)-2-(N-formimidoylpyrrolidin-3-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

413 (1R,5S,6S)-2-[(2R,4S)-2-(2-azetidinon-4-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

415 (1R,5S,6S)-2-[(2R,4S)-2-(2-azetidinon-3-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-methyl-1-carbapen-2-em-3-carboxylic acid,

417 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2R,4S)-2-(2-pyrrolidon-5-ylmethyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

419 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2R,4S)-2-(2-pyrrolidon-3-ylmethyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

421 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2R,4S)-2-(2-pyrrolidon-4-ylmethyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid,

441 (1R,5S,6S)-2-[(2R,4S)-2-(2-carbamoylpyrrolidin-4-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

450 (1R,5S,6S)-2-[(2R,4S)-2-(N,N-dimethyl-3-pyrrolidiniomethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

466 (1R,5S,6S)-2-[(2R,4S)-2-(3-amino-2-pyrrolidon-4-ylmethyl)pyrrolidin-4-ylthiol-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid,

476 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2R,4S)-2-(3-oxopiperazin-5-ylmethyl)pyrrolidin-4-ylthio]- 1-carbapen-2-em-3-carboxylic acid, and

498 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-2-[(2R,4S)-2-[N-(2-hydroxyethyl)pyrrolidin-3-ylmethyl]pyrrolidin-4-ylthio]-1-methyl-1-carbapen-2-em-3-carboxylic acid.

Especially preferred are compounds No. 142 i.e. (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(pyrrolidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid and No. 152 i.e. (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(piperidin-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid.

Now, the process for producing the compound of the present invention will be described.

An activating reagent is reacted to a compound of the formula: ##STR1123## wherein R is a hydrogen atom or a methyl group, R¹³ is hydrogen atom or a hydroxyl-protecting group, and R¹⁴ is a hydrogen atom or a carboxyl-protecting group in an inert organic solvent in the presence of a base to form a reactive derivative of the formula (II'): ##STR1124## wherein R, R¹³ and R¹⁴ are as defined above, and Y is a leaving group.

The inert organic solvent to be used for the reaction may, for example, be diethyl ether, tetrahydrofuran, dioxane, benzene, toluene, chlorobenzene, methylene chloride, chloroform, carbon tetrachloride, dichloroethane, trichloroethylene, acetone, ethyl acetate, acetonitrile, N,N-dimethylformamide, hexamethylphosphoric triamide-or a mixture of such solvents. Particularly preferred are acetonitrile and benzene.

The base to be used for the reaction may, for example, be a tertiary aliphatic amine such as trimethylamine, triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, N-methylpyrrolidine, N-methylpiperidine, N,N-dimethylaniline, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) or 1,5-diazabicyclo[4.3.0]non-5-ene (DBN); or an aromatic amine such as pyridine, 4-dimethylaminopyridine, picoline, lutidine, quinoline or isoquinoline. Particularly preferred are N,N-diisopropylethylamine and triethylamine.

The activating reagent to be used for the reaction may, for example, be an acid anhydride such as trifluoroacetic anhydride, methanesulfonic anhydride, trifluoromethanesulfonic anhydride or p-toluenesulfonic anhydride; or an acid chloride such as methanesulfonyl chloride, p-toluenesulfonyl chloride or diphenyl chlorophosphate. Particularly preferred is diphenyl chlorophosphate.

In the formula (II'), Y is a leaving group such as a trifluoroacetoxy group, a methanesulfonyloxy group, a trifluoromethanesulfonyloxy group, a p-toluenesulfonyloxy group or a diphenoxyphosphoryloxy group. Particularly preferred is a diphenoxyphosphoryloxy group.

For the reaction, from 1 to 3 mols, preferably from 1 to 1.5 mols, of the base and from 1to 1.2 mols of the activating reagent are used per mol of the compound of the formula (II).

The reaction is conducted usually within a temperature range of from -40° to 50° C., preferably from -20° to 20° C., and usually completed quantitatively in from 0.5 to 3 hours.

After completion of the reaction, the reaction product is treated in accordance with a usual method to obtain the reactive derivative (II') of the compound of the formula (II) quantitatively.

The reaction of the reactive derivative of the formula (II') with a compound of the formula: ##STR1125## wherein R¹⁵ is a hydrogen atom or an imino-protecting group, each of R²⁰ and R³⁰ which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group which may be protected, a formimidoyl group which may be protected, an acetoimidoyl group which may be protected, --COOR⁴⁰, --CON(R⁵⁰)R⁶⁰, --N(R⁵⁰)R⁶⁰, --CH₂ COOR⁴⁰, --CH₂ N(R⁵⁰)R⁶⁰ or --CH₂ CON(R⁵⁰)R⁶⁰ (wherein R⁴⁰ is a hydrogen atom, a lower alkyl group or a carboxyl-protecting group, and each of R⁵⁰ and R⁶⁰ which may be the same or different, is a hydrogen atom, a lower alkyl group, an amino-protecting group or an imino-protecting group, or R⁵⁰ and R⁶⁰ form together with the adjacent nitrogen atom a heterocyclic group selected from the group consisting of an aziridinyl group, an azetidinyl group, a pyrrolidinyl group and a piperidyl group), B is ═NR⁷⁰, ═N(R⁷⁰)R⁸⁰, --CON(R⁷⁰ )--, --CON(R⁷⁰)CO--, --CON(R⁷⁰)CON(R⁸⁰)--, --N(R⁷⁰)CO(CH₂)_(s) N(R⁸⁰)--, --N(R⁷⁰)CO(CH₂)_(s) CON(R⁸⁰)--, --CON(R⁷⁰)N(R⁸⁰)-- or --N(R⁷⁰)(CH₂)_(s) N(R⁸⁰)-- {wherein each of R⁷⁰ and R⁸⁰ which may be the same or different is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group which may be protected, a formimidoyl group which may be protected, an acetoimidoyl group which may be protected, an imino-protecting group, --COOR⁴⁰, --CON(R⁵⁰)R⁶⁰ , --N(R⁵⁰)R⁶⁰, --CH₂ COOR⁴⁰, --CH₂ N(R⁵⁰)R⁶⁰ or --CH₂ CON(R⁵⁰)R⁶⁰ (wherein R⁴⁰, R⁵⁰ and R⁶⁰ are as defined above), and s is an integer of from 1 to 3), and p, q and r are as defined above, is conducted using the above mentioned inert organic solvent and base to form a compound of the formula: ##STR1126## wherein R, R¹³, R¹⁴, R¹⁵, R²⁰, R³⁰, B, p, q and r are as defined above.

The reaction is conducted using from 1 to 2 mols, preferably from 1 to 15 mols, of the base and from 1 to 1.2 mols of the compound of the formula (III), per mol of the reactive derivative of the formula (II'). The reaction is conducted usually within a temperature range of from -40° to 50° C., preferably from -20° to 20° C., and the reaction is completed usually in from 0.5 to 3 hours.

Further, the compound of the formula (IV) can be prepared in one step from the compound of the formula (II). Namely, without isolating the reactive derivative of the formula (II') prepared from the compound of the formula (II), the compound of the formula (III) is reacted thereto in the same reaction system to prepare the compound of the formula (IV) efficiently. To conduct the production in one step, from 2 to 4 mols, preferably from 2.5 to 3.5 mols, of the base is employed per mol of the compound of the formula (II).

After completion of the reaction, usual treatment is conducted to obtain a crude product of the formula (IV), which may be subjected to a reaction for removing a protecting group without purification. However, it is preferred to purify the crude product (IV) by crystallization or by column chromatography by means of e.g. silica gel.

From the compound of the formula (IV) thus obtained, a compound of the formula (I) can be obtained, if necessary, by conducting a reaction for removing a protecting group for a hydroxyl group, an amino or imino group and a carboxyl group.

For the removal of the protecting groups, the method varies depending upon the type of the protecting groups. However, the removal can be conducted in accordance with conventional methods, for example, by solvolysis, by chemical reduction or by hydrogenation.

For example, when in the above formula (IV), the protecting group for the hydroxyl group and/or for the amino or imino group is an aralkyloxycarbonyl group such as a benzyloxycarbonyl group-or a p-nitrobenzyloxycarbonyl group, and the protecting group for the carboxyl group is an aralkyl group such as a benzyl group, a p-nitrobenzyl group or a benzhydryl group, such protecting groups can be removed by catalytic hydrogenation by means of a platinum catalyst such as platinum oxide, platinum wire or platinum black, or a palladium catalyst such as palladium black, palladium oxide, palladium-carbon or palladium hydroxide-carbon.

As a solvent to be used for such a catalytic hydrogenation reaction, methanol, ethanol, tetrahydrofuran, dioxane, acetic acid or a solvent mixture of such an organic solvent with water or with a buffer solution of e.g. a phosphate, may be used.

The reaction can be completed in from 0.5 to 4 hours at a temperature within a range of from 0° to 50° C. under hydrogen gas stream of from 1 to 4 atm.

When in the above formula (IV), the protecting group for the hydroxyl group and/or the amino or imino group is an allyloxycarbonyl group, and the protecting group for the carboxyl group is an allyl group, such protecting groups can be removed by reacting an organo-soluble palladium complex catalyst in an inert organic solvent containing an allyl group-capturing agent (method by W. McCombie et al., J. Org. Chem., vol. 47, p. 587-590 (1982) and method by F. Guibe, the same literature, vol. 52, p. 4,984-4,993 (1987)).

The solvent useful for the reaction includes, for example, water, acetone, diethyl ether, tetrahydrofuran, dioxane, ethyl acetate, acetonitrile, methylene chloride, chloroform and a solvent mixture thereof.

The palladium compound complex useful for this reaction includes, for example, palladium-carbon, palladium hydroxide-carbon, palladium(II) chloride, palladium(II) acetate, tetrakis(triphenylphosphine)palladium (O), tetrakis(triphenoxyphosphine)palladium (O), tetrakis(triethoxyphosphine)palladium (O), bis[ethylenebis(diphenylphosphine)]palladium (O), tetrakis[tri(2-furyl)phosphine]palladium (O), bis(triphenylphosphine)palladium(II) chloride and bis(triphenylphosphine)palladium(II) acetate.

The allyl group-capturing agent may, for example, be dimedone, formic acid, acetic acid, ammonium formate, sodium formate, sodium 2-ethylhexanoate, potassium 2-ethylhexanoate, pyrrolidine, piperidine and tributyltin hydride.

The reaction is conducted usually within a temperature range of from -10° to 50° C., preferably from 0 to 30° C. using from 0.01 to 0.5 mol of the catalyst and from 1 to 6 mols of the nucleophilic agent relative to 1 mol of the compound of the formula (IV), and the reaction is completed usually in from 0.5 to 3 hours.

Further, when in the above formula (IV), the protecting group for the hydroxyl group and/or the amino or imino group is an o-nitrobenzyloxycarbonyl group, and the protecting group for the carboxyl group is an o-nitrobenzyl group, such protecting groups can be removed by a photo reaction (method by Amit et al., J. Org. Chem., vol. 39, p. 192-196 (1974)).

After completion of the reactions for removing the protecting groups, the compound of the formula (I) can be isolated by usual treatment such as column chromatography using silica gel or adsorptive resin, freeze-drying or crystallization.

Further, when the protecting group for the carboxyl group at the 3-position of the compound of the (IV) is a lower alkanoyloxyalkyl group such as an acetoxymethyl group or a pivaloyloxymethyl group, a methoxymethyl group, an indanyl group, or a phthalidyl group, such an ester will be physiologically hydrolyzed in vivo. Therefore, such a compound can directly administered to a human being or to an animal without preliminarily removing the protecting group.

The compound of the formula (I) can be converted to a pharmaceutically acceptable salt or ester by a conventional method.

The starting material of the formula (II) can be prepared, for example, by a method by Salzmann et when R¹ is a hydrogen atom (J. Am. Chem. Soc., vol. 102, p.6161-6163 (1981)) or by a method by Shih et al. when R¹ is a methyl group (Heterocycles, vol. 21, p.29-40 (1984)).

The starting material of the formula (III) can be synthesized by the following method.

The hydroxyl group of the compound 1 is activated by a usual method, and a thioacetate such as potassium thioacetate is reacted thereto to convert it to an acetylthio derivative 3, followed by alkali or acid hydrolysis to obtain a thiol derivative of the formula (III). ##STR1127##

In the above formulas, R¹⁶ is a hydrogen atom or a hydroxyl-protecting group, X is a leaving group selected from the group consisting of a chlorine atom, a bromine atom, an iodine atom, a trifluoroacetoxy group, a methanesulfonyloxy group, a trifluoromethanesuifonyloxy group and a p-toluenesulfonyloxy group, Ac is an acetyl group, and R¹⁵, R²⁰, R³⁰, B, p, q and r are as defined above.

A group of compounds having the formula 1 can be prepared in accordance with the methods described in the Reference Examples.

The compounds of the present invention exhibit strong antibacterial activities against various gram positive bacteria and gram negative bacteria.

To demonstrate the usefulness of the compounds of the present invention, the in vitro antibacterial activities against bacteria were measured by the following agar plate dilution method (standard method by Japan Chemotherapy Society, Chemotherapy, vol. 29, p. 76-79 (1981)). One platinum loopful of each test microorganism incubated overnight in Mueller Hinton broth, was inoculated to Mueller Hinton agar (inoculum size: 10⁶ CFU/ml). Such culture media contained antibacterial agents in various concentrations. After incubation at 37° C. for 16 hours, the minimum inhibitory concentrations (MIC: μg/ml) were measured.

The results of the antibacterial activities of the compounds of the present invention are shown in Table 1.

                                      TABLE 1                                      __________________________________________________________________________     Minimum Inhibitory Concentration (MIC: μg/ml)                                       Example 11  Example 12                                                                           Example 13                                           Test    Diastereo-                                                                           Diastereo-                                                                           Diastereo-                                                                           Diastereo-                                                                           Diastereo-                                     microorganism                                                                          mer A mer B mer A mer A mer B Example 43                                                                           Example 49                                                                           Meropenem                                                                             Imipenem              __________________________________________________________________________     P. aeruginosa                                                                          0.1   0.2   --    0.2   0.2   0.1   0.2   0.39   1.56                  MB5000                                                                         P. aeruginosa                                                                          0.78  1.56  1.56  0.78  1.56  0.39  0.78  6.25   3.13                  MB5002                                                                         P. aeruginosa                                                                          0.78  1.56  1.56  1.56  --    1.56  1.56  3.13   6.25                  AKR17*                                                                         __________________________________________________________________________      *lactamase producing microorganism                                       

The antibacterial activities of the compounds of the present invention described in the Examples, as representative examples of the compound of the present invention, were measured by a disc diffusion test by the method of Bauer et al. (Amer. J. Clin. Pathol., vol. 45, p. 493 (1966)). Thienamycin or imipenem was used as the internal standard.

MIC of each test compound was calculated from the diameter of the inhibition ring formed by the disc containing the test compound by using the calculation formula reported by Humphrey and Lightbown (J. Gen. Microbiol., vol. 7, p. 129 (1952)). For each microorganism, a geometrical average of MIC was obtained, and the activity ratio to thienamycin was calculated.

The antibacterial activities are represented by the ratio to thienamycin (=1.0), whereby the larger the numerical value, the higher the activities.

The DHP-I susceptibility was quantitatively analyzed by the method by Kropp et al., Antimicrob. Agents Chemother., vol. 22, p. 62-70 (1982), whereby the smaller the numerical value representing the ratio to imipenem (=1.0), the higher the stability. The antibacterial potency and the DHP-I susceptibility of the compounds of the present invention were compared with imipenem and meropenem. The results are .shown in Table 2.

                                      TABLE 2                                      __________________________________________________________________________     Relative antibacterial potency to thienamycin and DHP-I susceptibility                Example 11                                                                     Diastereo-                                                                           Diastereo-                                                               mer A mer B Example 13                                                                           Example 32                                                                           Example 34                                                                           Example 49                                                                           Meropenem                                                                             Imipenem                     __________________________________________________________________________     Meth-R 15.2  12.3  --    --    22.7  13.6  3.22    2.57                        S. aureus                                                                      THM-R  12.7  11.5  11.5  19.5  21.6  13.8  6.9    2.0                          P. aeruginosa                                                                  DHP-I  <0.05 <0.05 <0.05 <0.05 0.07  0.10  0.12   1.0                          susceptibility                                                                 __________________________________________________________________________

The compounds of the present invention have excellent antibacterial activities against various gram positive bacteria and gram negative bacteria and are useful as antibacterial agents for the treatment and prevention of the human infectious diseases caused by such bacteria. Typical pathogens sensitive to the antibacterial agents of the present invention include, for example, species of genus Staphylococcus, genus Enterococcus, genus Escherichia, genus Enterobacter, genus Klebsiella, genus Serratia, genus Proteus and genus Pseudomonas. The compounds of the present invention exhibit excellent antibacterial activities particularly against Methicillin resistant Staphylococcus aureus and against thienamycin resistant Pseudomonas aeruqinosa.

The compounds of the present invention are very stable against DHP-I although the stability varies depending upon the individual compounds, and they are excellent also in the physicochemical stability and in the solubility in water.

The compounds of the present invention may be used in the form of drug formulations suitable for non-oral administration, oral administration or external administration, by mixing them with carriers of solid or liquid excipients known in this field. The main administration route is non-oral (intravenous or intramuscular injection) administration by injection or local administration. Drug formulations include liquid formulations such as injection solutions, syrups or emulsions, solid formulations such as tablets, capsules or granules, and external application formulations such as ointments or suppositories. These formulations may contain additives such as a base, an assisting agent, a stabilizer, a wetting agent, an emulsifier, an absorption-promoting agent, a surfactant, etc. which are commonly employed, as the case requires.

The additives include, for example, distilled water for injection, Ringer's solution, glucose, sucrose syrup, gelatin, edible oil, cacao butter, ethylene glycol, sucrose, corn starch, magnesium stearate and talc.

The dose varies depending upon the condition of the patient, the weight, the age, the sex, the type of formulation, the number of administration times, etc. Usually, however, a preferred daily dose of the active ingredient to an adult is from about 5 to 50 mg/kg, and a preferred daily dose to a child is within a range of from about 5 to 25 mg/kg, which is preferably administered once a day or in a few times a day.

The compound of the present invention may be administered in combination with a DHP-I inhibiting agent such as cilastatin [sodium (Z)-7-(L-amino-2-carboxyethylthio)-2-(2,2-dimethylcyclopropanecarboxamide)-2-heptenoate](Japanese Unexamined Patent Publication No. 81518/1981; European Patent No. 28,778; J. Med. Chem., vol. 30, p. 1074 (1987)).

Now, the present invention will be described in further detail with reference to Examples and Reference Examples. However, it should be understood that the present invention is by no means restricted by such specific Examples.

In the thin layer chromatography in the Examples and Reference Examples, silica gel 60F₂₄₅ (Merck) was used as the plate, and an ultraviolet detector was used as a detecting device. As the silica gel for the column, Wakogel™ C-300 (Wako Junyaku) was used, and as the silica gel for reversed phase column, LC-SORB™ SP-B-ODS (Chemco) or YMC-GEL^(TM) ODS-AQ 120-550 (Yamamura Chemical Laboratories) was used. As the high pressure liquid chromatograph, JASCO 800 series (Nippon Bunko) was used. When the NMR spectrum was measured using a dimethyl sulfoxide-d₆ or chloroform-d solution, tetramethylsilane (TMS) was used as the internal standard, and when measured using a deuterium oxide solution, 2,2'-dimethyl-2-silapentane-5-sulfonate (DSS) was used as the internal standard, and the measurement was conducted by means of XL-200 (200 MHz;Varian) model spectrometer. All δ values are shown by ppm.

The meanings of the abbreviations used for the NMR measurement are as follows:

s: singlet

d: doublet

t: triplet

q: quartet

ABq: AB-type quartet

dd: double doublet

m: multiplet

br: broad

J: coupling constant

Hz: hertz

DMSO-d₆ : dimethyl sulfoxide-d₆

CDCl₃ : chloroform-d

CD₃ OD: methanol-d₄

D₂ O: deuterium oxide

The meanings of the abbreviations used in the reaction formulas are as follows:

Ac: acetyl group

All: allyl group

Alloc: allyloxycarbonyl group

Boc: tert-butoxycarbonyl group

Bzl: benzyl group

Et: ethyl group

Me: methyl group

Ms: methanesulfonyl group

PNB: p-nitrobenzyl group

PNZ: p-nitrobenzyloxycarbonyl group

TBDMS: tert-butyldimethylsilyl group

Tr: trityl group

EXAMPLE Sodium (5R,6S)-6-[(R)-1-hydroxyethyl]-2-[(2S,4S)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate ##STR1128##

To a solution of p-nitrobenzyl (5R,6S)-2-diphenoxy phosphoryloxy-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (300 mg, 0.55 mmol) in acetonitrile (15 ml) were dropwise added in a nitrogen stream under cooling with ice a solution of (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine (205 mg, 0.56 mmol, Compound of Reference Example 1-9) in acetonitrile (6 ml) and then N,N-diisopropylethylamine (0.10 ml, 0.57 mmol). The mixture was stirred at 0° C. for 7 hours. Then, ethyl acetate (70 ml) was added to the reaction solution. The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate) to obtain p-nitrobenzyl (5R,6S)-6-[ (R)-1-hydroxyethyl]-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (313 mg, yield: 86.2%).

1R(KBr)cm⁻¹ : 1780, 1700, 1520, 1350

NMR(CDCl₃) δ: 1.37(3H,d,J=6Hz),5.24(3H,m),5.52(1H,d,J=14Hz),7.33 (2H,d,J=9Hz),7.46(2H,d,J=9Hz),8.24(2H,d,J=9Hz),8.26 (2H,d,J=9Hz) ##STR1129##

10% palladium-carbon catalyst (150 mg preliminarily stirred and activated with a 0.1 M sodium 3-morpholinopropanesulfonate buffer solution in a hydrogen stream for one hour) was added to a solution of the compound obtained by the above reaction (300 mg, 0.45 mmol) in a mixture of tetrahydrofuran (10 ml) and a 0.1M sodium 3-morpholinopropanesulfonate buffer solution (10 ml). This mixture was stirred in a hydrogen stream at room temperature for two hours. The catalyst was filtered off from the reaction mixture, and the filtrate was washed with ethyl acetate (20 ml), and insoluble matters in the aqueous layer were filtered off. The obtained filtrate was concentrated to a volume of about 15 ml. The residue was subjected to reversed phase column chromatography (LC-SORB™ SP-B-ODS, 10% methanol aqueous solution). The desired fraction was concentrated and freeze-dried to obtain the above identified compound (70 mg, yield: 38.4%).

1R(KBr)cm⁻¹ : 1760, 1680, 1590, 1390

NMR(D₂ O) δ: 1.26(3H,d,J=7Hz),1.54(1H,m),2.28(1H,m),3.86(1H,m), 4.21(2H,m)

HPLC:

Column: YMC™-Pack ODS-AQ, 5μ, 4.6φ×150 mm

Eluent: 0.01M Phosphate buffer (pH 6.5)-Methanol (80:20)

Flow rate: 1.0 ml/min

Temperature: 40° C.

Detector: 290 nm

Retention time: 2.23 min

EXAMPLE 2 Sodium (5R,6S)-6-[(R)-1-hydroxyethyl]-2-[(2R,4S)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate ##STR1130##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (5R,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (300 mg, 0.55 mmol) and (2R,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine (200 mg, 0.55 mmol, compound of Reference Example 2) to obtain p-nitrobenzyl (5R,6S)-6-[(R)-1-hydroxyethyl]-2-(2R,4S)-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (127 mg, yield: 35%).

1R(KBr)cm⁻¹ : 1790, 1710, 1520, 1350

NMR(CDCl₃) δ: 1.36(3H,d,J=6Hz),5.25(3H,m),5.52(1H,d,J=14Hz),7.54 (2H,d,J=9Hz),7.66(2H,d,J=9Hz),8.23(2H,d,J=9Hz), 8.26(2H,d,J=9Hz) ##STR1131##

10% palladium-carbon catalyst (60 mg) was added to a solution of the compound obtained by the above reaction (127 mg, 0.19 mmol) in a mixture of tetrahydrofuran (10 ml) and a 0.1M sodium 3-morpholinopropanesulfonate buffer solution (10 ml). This mixture was stirred in a hydrogen stream of 2.9 atm at room temperature for 1.5 hours. The catalyst was filtered off from the reaction mixture, and the filtrate was washed with ethyl acetate (20 ml), and insoluble matters in the aqueous layer were filtered off. The obtained filtrate was subjected to reversed phase column chromatography (LC-SORB^(TM) SP-B-ODS, 10% methanol aqueous solution), then concentrated and freeze-dried to obtain the above identified compound (5.5 mg, yield: 7.1%).

1R(KBr)cm⁻ : 1780, 1600, 1270

NMR(D₂ O) δ:

1.28(3H,d,J=8Hz),2.54(3H,m)

HPLC (the same condition as in Example 1)

Retention time: 5.54 min

EXAMPLE 3 Sodium (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthiol]-1-carbapen-2-em-3-carboxylate ##STR1132##

The same procedure as in Example 1-1 was conducted by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (230 mg, 0.41 mmol) and (2S,4S)-4-mercapto-N-(P-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine (140 mg, 0.3 8 mmol, Compound of Reference Example 1-9) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1hydroxyethyl]-1-methyl-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl ) pyrrolidin-4-ylthio]-1-carbapen-2-em-3 -carboxylate (223 mg, yield: 80.6%).

1R(KBr)cm⁻¹ : 1770, 1700, 1520, 1340

NMR(CDCl₃) δ: 1.28(3H,d,J=8Hz),1.33(3H,d,J=7Hz),5.24(2H,m),5.31 and 5.52(2H,ABq,J=14Hz),7.53(2H,d,J=8Hz),7.66(2H,d, J=8Hz),8.20(2H,d,J=8Hz),8.22(2H,d,J=8Hz) ##STR1133##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (223 mg, 0.33 mmol) to obtain the above identified compound (73 mg, yield: 53.2%, diastereomers A and B 44:56).

1R(KBr )cm⁻¹ : 1760, 1680, 1590, 1390

NMR(D₂ O) δ: 1.23(3H,d,J=8Hz),1.30(3H,d,J=7Hz) ,1.77(1H,m), 2.27-2.42(1H,m) ,4.06(1H,m) ,4.26(2H,m)

HPLC:

Column: INERTSIL™ ODS-2, 5μ, 4.6φ×250 mm

Eluent: 0.01M Phosphate buffer (pH7.0)-Methanol (90:10)

Flow rate: 1.0 ml/min

Temperature: 40° C.

Detector: 254 nm

Retention time: 13.2 min, 14.6 min (44:56)

EXAMPLE 4 Sodium (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2R,4S)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate ##STR1134##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (230 mg, 0.41 mmol) and (2R,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine (140 mg, 0.38 mmol, compound of Reference Example 2) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2R,4S)-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (174 mg, yield: 62.9%).

1R(KBr)cm⁻¹ : 1780, 1700, 1520, 1340

NMR(CDCl₃) δ: 1.27(3H,d,J=8Hz),l.36(3H,d,J=6Hz),5.25(3H,m),5.53(1H, d,J=14Hz),7.54(2H,d,J=9Hz),7.67(2H,d,J=9Hz),8.24(2H, d,J=9Hz),8.26(2H,d,J=9Hz) ##STR1135##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (174 mg, 0.26 mmol) to obtain the above identified compound (22 mg, yield: 20.6%).

1R(KBr)cm⁻¹ : 1780, 1600, 1380, 1300

NMR(D₂ O) δ: 1.24(3H,d,J=8Hz),1.31(3H,d,J=7Hz),1.98-2.98 (3H,m),3.44(3H,m),3.74(3H,m)

HPLC (the same condition as in Example 1)

Retention time: 7.46 min

EXAMPLE 5 Sodium (1R,5S,6S)-2-[(2S,4S)-2-azetidinon-4-yl)pyrrolidin-4-ylthio)-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer B ##STR1136##

To a solution of p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl)]-1-methyl-1-carbapen-2-em-3-carboxylate (110 mg, 0.195 mmol) in acetonitrile (5ml) was added in a nitrogen stream at -10° C. a solution of (2S,4S)-2-(2-azetidinon-4-yl)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer B (67 mg, 0.191 mmol, compound of Reference Example 3) in acetonitrile (5ml), and the N,N-diisopropylamine (34 μl, 0.21 mmol) was dropwise added thereto. The mixture was stirred overnight at 4° C. Then, ethyl acetate (50 ml) was added to the reaction solution. This mixture was washed with water and a saturated sodium chloride aqueous solution. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 3% methanol-chloroform) to obtain p-nitrobenzyl (1R,5S,6S)-2-[(2S,4S)-2-(2-azetidinon-4-yl)-N-(P-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer B (88.5 mg, yield: 68.2%).

1R(KBr)cm⁻¹ : 1760, 1700, 1520, 1340

NMR(CDCl₃) δ: 1.28(3H,d,J=7Hz),1.35(3H,d,J=6Hz),1.67(1H,m), 2.50-2.74(3H,m),3.08(1H,dd,J=15,5Hz),3.24-3.45 (3H,m),3.70(2H,m),3.96-4.33(4H,m),5.23(3H,m), 5.52(1H,d,J=13Hz),7.52(2H,d,J=8Hz),7.65 (2H,d,J=8Hz),8.22(2H,d,J=8Hz),8.24(2H,d,J=8Hz) ##STR1137##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (88 mg, 0.13 mmol) to obtain the above identified compound (18 mg, yield: 33.7%).

1R(KBr)cm⁻¹ : 1750, 1590, 1390

NMR(D₂ O) δ: 1.26(3H,d,J=7Hz),1.31(3H,d,J=6Hz),1.65(1H,m),2.69(1H, dd,J=16,8Hz),2.73(1H,br d,J=16Hz),3.20-3.53(4H,m), 3.61(1H,dd,J=12,6Hz),3.75(1H,q,J=8Hz),3.85-4.40(4H,m)

HPLC (the same condition as in Example 1)

Retention time: 3.8 min

EXAMPLE 6 Sodium (1R,5S,6S)-2-[(2S,4S)-2-(2-azetidinon-4-yl)pyrrolidin-4-ylthio)-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer A ##STR1138##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (153 mg, 0.27 mmol) and (2S,4S)-2-(2-azetidinon-4-yl)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer A (90 mg, 0.26 mmol, compound of Reference Example 4) to obtain p-nitrobenzyl (1R,5S,6S)-2-[(2S,4S)-2-(2-azetidinon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer A (128 mg, yield: 70.9%).

1R(KBr)cm³¹ 1 : 1760, 1700, 1520, 1340

NMR(CDCl₃) δ: 1.28(3H,d,J=7Hz),1.34(3H,d,J=6Hz),1.91(2H,m), 2.44-3.10(3H,m),3.17-3.46(2H,m),3.66(1H,m), 4.00-4.36(5H,m),5.24(3H,m),5.50(1H,d,J=14Hz), 7.52(2H,d,J=8Hz),7.65(2H,d,J=8Hz),8.21(2H,d,J=8Hz), 8.23(2H,d,J=8Hz) ##STR1139##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (128 mg, 0.19 mmol) to obtain the above identified compound 28.5 mg, yield: 36.7%).

1R(KBr)cm⁻¹ : 1750, 1590, 1390

NMR(D₂ O) δ: 1.25(3H,d,J=7Hz),1.32(3H,d,J=6Hz),1.83(1H,m),2.75(1H, dd,J=15,8Hz),2.88(1H,dd,J=15,2Hz),3.18-3.55(4H,m), 3.68(1H,dd,J=12,6Hz),3.86-4.40(5H,m)

HPLC (the same condition as in Example 1)

Retention time: 3.46 min

EXAMPLE 7 (5R,6S)-6-[(R)-1-hydroxyethyl]-2-[(2R,4S)-2-(z-pyrrolidon-3-ylmethyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid ##STR1140##

N,N-diisopropylethylamine (0.13 ml, 0.76 mmol) was dropwise added to a solution of allyl (5R,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (369 mg, 0.76 mmol) and (2R,4S)-N-allyloxycarbonyl-4-mercapto-2-(2-pyrrolidon-3-ylmethyl)pyrrolidine (216 mg, 0.76 mmol) in acetonitrile (5.6 ml) under cooling with ice. The reaction mixture solution was stirred at the same temperature for one hour and further stirred at 5° C. for 16 hours. Ethyl acetate (60 ml) was added to the reaction solution. The mixture was washed sequentially with a saturated sodium hydrogen carbonate aqueous solution and a saturated sodium chloride aqueous solution, then dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel flash column chromatography (Wakogel™ C-300, 40 ml, acetone-ethyl acetate 2:3). The fraction containing the desired product was concentrated to obtain allyl (5R,6S)-2-[(2R,4S)-N-allyloxycarbonyl-2-(2-pyrrolidon-3-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (303 mg, yield: 76.7%) in the form of foam.

NMR(CDCl₃) δ: 1.35(3H,d,J=6Hz),1.7-2.2(4H,m),2.34(2H,m),2.61(1H,m), 3.1-3.4(6H,m),3.55(1H,m),4.0-4.3(4H,m),4.6-4.9(4H,m), 5.2-5.5(4H,m),5.78(1H,br s),6.0(2H,m) ##STR1141##

Water (52 μl) was added to a solution of the compound obtained by the above reaction (300 mg, 0.58 mmol) in methylene chloride (6 ml). This mixture was deaerated. Bis(triphenylphosphine)palladium(II) chloride (8 mg, 0.011 mmol) and tributyltin hydride (0.466 ml, 1.73 mmol) were added to this solution mixture under cooling with ice. This solution mixture was stirred at the same temperature for 5 minutes and further at room temperature for 30 minutes. Water (40 ml) was added to the reaction solution. The aqueous layer was washed with chloroform (20 ml, twice), and then the remaining organic solvent was removed under reduced pressure. Active carbon (50 mg) was added thereto. This mixture was stirred for 30 minutes and then filtered. The filtrate was concentrated to 700 mg, and ethanol (1.4 ml) was added thereto at room temperature. This solution mixture was left to stand at the same temperature for 30 minutes to form precipitate. To this suspension, ethanol (1.4 ml) was dropwise added under stirring over a period of one hour. Then, the suspension was stirred at room temperature for 30 minutes and further at 5° C. for 16 hours. The precipitate was collected by filtration and washed sequentially within a water-ethanol (1:4) mixture (1.3 ml, twice) and acetone (2 ml) and then dried under reduced pressure for two hours to obtain the above identified compound (159 mg, yield: 69.6%).

IR(KBr)cm⁻¹ : 1740, 1690, 1600, 1380

NMR(D₂ O) δ: 1.26(3H,d,J=8Hz),1.7-2.0(3H,m),1.18(1H,m),1.38(1H,m), 2.6-2.8(2H,m),3.2(2H,d,J=9Hz),3.3-3.4(4H,m),3.7-3.9 (2H,m),4.0(1H,m),4.1-4.3(2H,m)

EXAMPLE 8 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2R,4S)-2-(2-pyrrolidon-3-ylmethyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid ##STR1142##

N,N-diisopropylethylamine (0.26 ml, 1.52 mmol) was dropwise added to a solution of allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (759 mg, 1.52 mmol) and (2R,4S)-N-allyloxycarbonyl-4-mercapto-2-(2-pyrrolidon-3ylmethyl)pyrrolidine (432 mg, 1.52 mmol) in acetonitrile (11 ml) at -40° C. The reaction solution mixture was stirred at the same temperature for 3 hours and further at 5° C. for 16 hours. Ethyl acetate (60 ml) was added to the reaction solution. This solution mixture was washed sequentially with a saturated sodium hydrogen carbonate aqueous solution and a saturated sodium chloride aqueous solution, then dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel flash column chromatography (Wakogel™ C-300, 40 ml, acetone-ethyl acetate 2:3), and the fraction containing the desired product was concentrated to obtain allyl (1R,5S,6S)-2-[(2R, 4S)-N-allyloxycarbonyl-2-(2-pyrrolidon-3-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (420 mg, yield: 51.8%) in the form of foam.

NMR(CDCl₃) δ: 1.26(3H,d,J=8Hz),1.35(3H,d,J=6Hz),1.7(1H,m),1.9-2.4 (5H,m),2.60(1H,m),3.2-3.4(5H,m),3.58(1H,m),3.9-4.3 (4H,m),4.5-4.9(4H,m),5.2-5.5(4H,m),5.8-6.1(3H,m) ##STR1143##

Water (71 μl) was added to a solution of the compound obtained by the above reaction (420 mg, 0.79 mmol) in methylene chloride (8.4 ml). This solution was deaerated. Bis(triphenylphosphine)palladium(II) chloride (11 mg, 0.016 mmol) and tributyltin hydride (0.635 ml, 2.36 mmol) were added to this solution mixture under cooling with ice. This solution mixture was stirred at the same temperature for 3 minutes and further at room temperature for 30 minutes. Water (40 ml) was added to the reaction solution, and the aqueous layer was washed with chloroform (20 ml, twice). Then, the remaining organic solvent was removed under reduced pressure. Active carbon (50 mg) was added thereto. This mixture was stirred for 30 minutes and then filtered. The filtrate was concentrated to 500 mg, and ethanol (1 ml) was added thereto at room temperature. This solution mixture was left to stand at the same temperature for one hour to form precipitate. To this suspension, ethanol (3.5 ml) was dropwise added under stirring over a period of one hour. This suspension was stirred at room temperature for 30 minutes and further at 5° C. for 16 hours. The precipitate was collected by filtration, then washed sequentially with a water-ethanol (1:9) mixture (1 ml, three times) and acetone (3 ml) and dried for two 10 hours under reduced pressure to obtain the above identified compound (232 mg, yield: 72.0%).

IR(KBr)cm⁻¹ : 1760, 1700, 1640, 1590, 1390

NMR(D₂ O) δ: 1.21(3H,d,J=8Hz),1.28(3H,d,J=6Hz),1.7-2.0(3H,m),2.20 (1H,m),2.40(1H,m),2.6-2.9(2H,m),3.3-3.5(5H,m),3.70 (1H,m),3.86(1H,m),4.0(1H,m),4.24(2H,m)

EXAMPLE 9 (1R,5S,6S)-2-[(2R,4S)-2-(2-azetidinon-3-ylmethyl)-pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid ##STR1144##

The same procedure as in Example 8-1 was carried out by using allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (331 mg, 0.66 mmol), (2R,4S)-N-allyloxycarbonyl-2-(2-azetidinon-3-ylmethyl)-4-mercaptopyrrolidine (180 mg, 0.66 mmol) and N,N-diisopropylethylamine (0.12 ml, 0.66 mmol) to obtain allyl (1R,5S,6S)-2-[(2R,4S)-N-allyloxycarbonyl-2-(2-azetidinon-3-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (125 mg, yield: 36.3%) in the form of foam.

NMR(CDCl₃) δ: 1.29(3H,d,J=7Hz),1.36(3H,d,J=6Hz),1.8(1H,m),2.0(1H,m),2.65(2H,m),3.1(1H,m),3.3(3H,m),3-48(1H,t, J=6Hz),3.6(1H,m),4.0(2H,m),4.25(2H,m),4-62(2H, br d, J=6Hz),4.8(2H,m),5.2-5.5(4H,m),5-66(1H, br s), 5.98(2H,m) ##STR1145##

The same protecting group removal reaction and post treatment as in Example 8-2 were carried out by using the compound obtained by the above reaction (125 mg, 0.24 mmol), bis(triphenylphosphine)palladium(II) chloride (3.4 mg, 0.0048 mmol), tributyltin hydride (0,194 ml, 0.722 mmol) and water (22 μl). The obtained aqueous layer was purified by reversed phase column chromatography (YMC.GEL™ ODS-AQ 120-S50, 50 ml, methanol-water 15:85), and the fraction containing the desired product was concentrated and freeze-dried to obtain the above identified compound (20 mg, yield: 21.0%).

IR(KBr)cm⁻¹ : 1740, 1600, 1390

NMR(D₂ O) δ: 1.18(3H,d,J=7Hz),1.26(3H,d,J=6Hz),1.3(1H,m),2.2 (3H,m),2.7(1H,m),3.1-3.7(7H,m),3.92(1H,m),4.1-4.3 (2H,m)

EXAMPLE 10 Potassium (1R,5S,6S)-6-[(R),l-hydroxyethyl.]-1-methyl-2-[(2S,4S)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthiol-1-carbapen-2-em-3-carboxylate diastereomers A and B ##STR1146##

The compound of Example 3 (100 mg) was subjected to Waters 600E (column: YMC™-Pack SH-365-5 S-5 120A ODS, eluent: 0.01 M potassium phosphate buffer (pH 7.0)-methanol 85:15, flow rate: 14 ml/min, detector: 290 nm, temperature: 26° C.) to obtain fractions of diastereomer A (retention time: 25.5 min) and diastereomer B (retention time: 30.2 min), respectively. The fraction containing diastereomer A was concentrated to about 15 ml, and the concentrated solution was subjected to reversed phase column chromatography (YMC™.GE_(L) ODS-AQ 120-S50, distilled water was passed to remove salts and then the desired product was eluted by a 20% methanol aqueous solution). The desired fraction was concentrated and freeze-dried to obtain diastereomer A (16 mg) of the above identified compound. In the same manner as above, diastereomer B (14.8 mg) was obtained.

Diastereomer A

1R(KBr)cm⁻¹ : 1760, 1680, 1590, 1390

NMR(D₂ O) δ: 1.22(3H,d,J=8Hz),1.30(3H,d,J=7Hz),1.67(1H,m),2.35 (1H,dd,J=8,17Hz),4.02(1H,m),4.25(2H,m)

Diastereomer B

1R(KBr)cm⁻¹ : 1760, 1680, 1590, 1390

NMR(D₂ O) δ: 1.22(3H,d,J=8Hz),1.30(3H,d,J=7Hz),1.72(1H,m),2.28 (1H,dd,J=8,17Hz),4.02(1H,m),4.26(2H,m)

EXAMPLE 11 (1R, 5S,6S)-6-[(R)-1-hydroxyethyl ]-1-methyl-2-[(2S,4S)-2-(pyrrolidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid diastereomers A and B ##STR1147##

The same procedure as in Example 1--1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (1.39 g, 2.34 mmol) and (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidine (1.24 g, 2.34 mmol, compound of Reference Example 5-6) to obtain p-nitrobenzyl (IR,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidin-4ylthio]-1-carbapen-2-em-3-carboxylate (1.59 g, yield: 77.7%).

1R(KBr)cm⁻¹ : 1770, 1700, 1610, 1520, 1400, 1350

NMR(CDCl₃) δ: 1.28(3H,d,J=7Hz),1.34(3H,d,J=6Hz),5.14-5.58 (6H,m),7.52(4H,br d,J=8Hz),7.65(2H,d,J=8Hz), 8.20(6H,br d,J=8Hz) ##STR1148##

The compound obtained by the above reaction (1.52 g, 1.74 mmol) was dissolved in a solution mixture comprising tetrahydrofuran (40 ml), ethanol (2 ml) and a 0.25M sodium 3-morpholinopropanesulfonate buffer solution (pH 7.0, 18 ml). Then, 10% palladium-carbon catalyst (750 mg) was added thereto. This mixture was stirred in a hydrogen stream of 3 atm at room temperature for two hours. The catalyst was filtered off from the reaction mixture, and the filtrate was washed with ethyl acetate (50 ml). Then, insoluble matters in the aqueous solution were filtered off. The aqueous layer thus obtained was subjected to reversed phase column chromatography (LC-SORB™ SP-B-ODS, 15% methanol aqueous solution →20% methanol aqueous solution), then concentrated and freeze-dried to obtain diastereomer A (97 mg, yield: 14.6%) and diastereomer B (110 mg, yield: 16.6%) of the above identified compound, respectively.

Diastereomer A

1R(KBr)cm⁻¹ : 1760, 1580, 1540, 1380

NMR(D₂ O) δ: 1.16(3H,d,J=8Hz),1.24(3H,d,J=7Hz),1.74(1H,m),2.20(1H,m),2.44(2H,m),3.73(1H,m),4.17(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 2.6 min

Diastereomer B

1R(KBr)cm⁻¹ : 1750, 1590, 1390

NMR(D₂ O) δ: 1.23(3H,d,J=8Hz),1.30(3H,d,J=7Hz),1.74(1H,m),2-23 (1H,m),2.50(2H,m),3.78(1H,m),4.24(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 3.6 min

EXAMPLE 12 (1R, 5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,45)-2-(N-methylpyrrolidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid diastereomers A and B ##STR1149##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (680 mg, 1.14 mmol) and (2S,4S)-4-mercapto-2-(N-methylpyrrolidin-3-yl)-N-(P-nitrobenzyloxycarbonyl)pyrrolidine trifluoromethane sulfonate (650 mg, 1.26 mmol, compound of Reference Examples 6-9) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-(N-methylpyrrolidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (603 mg, yield: 74.3%).

1R(KBr)cm⁻¹ : 1770, 1700, 1610, 1520

NMR(CDCl₃) δ: 1.37(3H,d,J=6Hz),1.42(3H,d,J=7Hz),5.24(4H,m),7.54 (2,d,J=8Hz),7.68(2H,d,J=8Hz ),8.25(2H,d,J=8Hz),8.27 (2H,d,J=8Hz) ##STR1150##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (290 mg, 0.41 mmol) to obtain diastereomer A (27 mg, yield: 16.7%) and diastereomer B (27 mg, yield: 16.7%) of the above identified compound, respectively.

Diastereomer A

1R(KBr)cm⁻¹ : 1760, 1590,

NMR(D₂ O) δ: 1.21(3H,d,J=8Hz),1.30(3H,d,J=7Hz),1.53(1H,m),1.92 (1H,m),2.94(3H,s),3.9t(1H,m),4.23(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 3.5 min

Diastereomer B

1R(KBr)cm⁻¹ : 1750, 1590, 1380

NMR(D₂ O) δ: 1.18(3H,d,J=8Hz),1.27(3H,d,J=7Hz),1.80(1H,m),2.24 (1H,m),2.88(3H,s),3.52(1H,m),3.74(1H,m),4.20(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 4.3 min

EXAMPLE 13 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2[(2S,4S)-2-(N,N-dimethyl-pyrrolidinio)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate diastereomers A and B ##STR1151##

Methyl iodide (0.13 ml, 2.09 mmol) was added to a solution of the compound obtained in Example 12-1 (300 mg, 0.42 mmol) in acetone (5 ml), and the mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the obtained residue was treated in the same manner as in Example 1-2 to obtain diastereomer A (17 mg, yield: 9.8%) and diastereomer B (29 mg, yield: 16.7%) of the above identified compound, respectively.

Diastereomer A

1R(KBr)cm⁻¹ : 1750, 1590, 1380

NMR(D₂ O) δ: 1.20(3H,d,J=8Hz),1.29(3H,d,J=7Hz),2.12(1H,m),2.44 (2H,m),3.17(3H,s),3.24(3H,s),3.77(2H,m),4.22(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 2.7 min

Diastereomer B

1R(KBr )cm³¹ 1 : 1750, 1590, 1380

NMR(D₂ O) δ: 1.22(3H,d,J=8Hz),1.30(3H,d,J=7Hz), 2.02 (1H,m), 2.52 (2H,m), 3.17(3H,s),3.25(3H,s), 3.78(2H,m), 4.22(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 3.5 min

EXAMPLE 14 Sodium (5R,6S)-6-[(R)-1-hydroxyethyl]-2-[(2S,4S),-2-(N-methyl-2-azetidinon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate ##STR1152##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (5R,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (235 mg, 0.40 mmol) and (2S,4S)-4-mercapto-2-(N-methyl-2-azetidinon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (149 mg, 0.41 mol, compound of Reference Example 7) to obtain p-nitrobenzyl (5R,6S)-6-[(R)-1-hydroxyethyl]-2-[(2S,4S)-2-(N-methyl-2-azetidinon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (265 mg, yield: 94.5%).

IR(KBr)cm⁻¹ : 1780, t740, 1700, 1520, 1340

NMR(CDCl₃) δ: 1.36(3H,d,J=6Hz),2.78(3H,s),5.26(3H,m),5.52 (1H,d,J=14Hz),7.54(2H,d,J=8Hz),7.66(2H,d,J=8Hz), 8.22(2H,d,J=8Hz),8.24(2H,d,J=8Hz) ##STR1153##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (265 mg, 0.37 mmol) to obtain diastereomer A (4 mg, yield: 2.7%), diastereomer B (25 mg, yield: 16.7%) and a mixture of diastereomers A and B (25 mg, yield: 16.7%) of the above identified compound.

Diastereomer A

IR(KBr)cm⁻¹ : 1750, 1590, 1390

NMR(D₂ O) δ: 1.27(3H,d,J=6Hz),1.66(1H,m),2.74(2H,m),2-90(3H,s), 3.10-3.47(5H,m),3.67(2H,m),4.22(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 2.68 min

Diastereomer B

IR(KBr)cm⁻¹ : 1750, 1590, 1390

NMR(D₂ O) δ: 1.28(3H,d,J=6Hz),1.89(1H,m),2.88(3H,s),2.92 (2H,m),3.16-3.48(5H,ml,3.75(1H,dd,J=12,6Hz), 4.24(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 2.85 min

EXAMPLE 15 Sodium (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methyl-2-azetidinon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate ##STR1154##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (240 mg, 0.40 mmol) and (2S,4S)-4-mercapto-2-(N-methyl-2-azetidinon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (149 mg, 0.41 mmol) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methyl-2-azetidinon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (236 mg, yield: 82.7%).

IR(KBr)cm⁻¹ : 1750, 1700, 1520, 1350

NMR(CDCl₃) δ: 1.28(3H,d,J=7Hz),1.36(3H,d,J=6Hz),2.78(3H,s), 5.26(3H,m),5.50(1H,d,J=14Hz),7.54(2H,d,J=8Hz), 7.67(2H,d,J=8Hz),8.22(2H,d,J=8Hz),8.24(2H,d,J=8Hz) ##STR1155##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (236 mg, 0.33 mmol) to obtain diastereomer A (25 mg, yield: 18.1%) and diastereomer B (39 mg, yield: 28.2%) of the above identified compound.

Diastereomer A

IR(KBr)cm⁻¹ : 1750, 1600,

NMR(D₂ O) δ: 1.22(3H,d,J=7Hz),1.30(3H,d,J=6Hz),1.78(1H,m),2.81 (2H,m),2.94(3H,s),3.20-3.55(5H,m),3.74 (1H,dd,J=12,6Hz),4.26(2H,m)HPLC (the same condition as in Example 1)

Retention time: 4.86 min

Diastereomer B

IR(KBr)cm⁻¹ : 1750, 1600,

NMR(D₂ O) δ: 1.24(3H,d,J=7Hz),1.30(3H,d,J=6Hz),1.88(1H,m),2.90 (3H,s),2.95(2H,m),3.18-3.34(5H,m),3.70 (1H,dd,J=12,6Hz),4.26(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 5.43 min

EXAMPLE 16 Sodium (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methyl-2,5-dioxopyrrolidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate diastereomer A ##STR1156##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate(500 mg, 0.84 mmol) and (2S,4S)-4-mercapto-2-(N-methyl-2,5-dioxopyrrolidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer A (330 mg, 0.84 mmol, compound of Reference Example 8) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methyl-2,5-dioxopyrrolidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate diastereomer A (311 mg, yield: 50.1%).

IR(KBr)cm⁻¹ : 1770, 1700, 1520, 1350

NMR(CDCl₃) δ: 1.28(3H,d,J=8Hz),1.34(3H,d,J=7Hz),2.95(3H,br s),3.68 (1H,m),4.57(1H,m),5.20(3H,m),5.52(1H,d,J=i4Hz),7.48 (2H,d,J=8Hz),7.66(2H,d,J=8Hz),8.22(4H,d,J=8Hz) ##STR1157##

The same procedure as in Example 1-2 was carried out by using the compound obtained in the above reaction (311 mg, 0.42 mmol) to obtain the above identified compound (76.8 mg, yield: 40.9%).

IR(KBr)cm⁻¹ : 1750, 1700, 1610, 1590

NMR(D₂ O) δ: 1.20(3H,d,J=7Hz ),1.26(3H,d,J=6Hz),1.95(1H,m), 2.96(3H,s),3.25-3.74(5H,m),4.00(2H,m),4.20(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 4.60 min

EXAMPLE 17 Sodium (1R,5S,6S)-6-[(R)-1-hydroxyethyl ]-1-methyl-2-[(2S,4S )-2-(N-methyl-2,5-dioxopyrrolidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate diastereomer B ##STR1158##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (220 mg, 0.37 mmol) and (2S,4S)-4-mercapto-2-(N-methyl-2,5-dioxopyrrolidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer B (140 mg, 0.36 mmol) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methyl-2,5-dioxopyrrolidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate diastereomer B (244 mg, yield: 89.4%).

IR(KBr)cm⁻¹ : 1780, 1700, 1520,

NMR(CDCl₃) δ: 1.28(3H,d,J=7Hz),1.34(3H,d,J=6Hz),1.62(1H,m),2.96 (3H,s),5.26(3H,m),5.50(1H,d ,J=l 4Hz),7.54 (2H,d,J=8Hz),7.67(2H,d,J=8Hz),8 22(4H,d,J=8Hz) ##STR1159##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (244 mg, 0,322 mmol) to obtain the above identified compound (35 mg, yield: 23.7%).

IR(KBr)cm⁻¹ : 1750, 1700, 1610, 1590

NMR(D₂ O) δ: 1.22(3H,d,J=7Hz),1.30(3H,d,J=6Hz),1.84(1H,m),2.66 (1H,dd,J=18,6Hz),2.84(1H,m),2.98(3H,s),3.12 (1H,dd,J=18Hz,9Hz),3.30-3.58(4H,m),3.72 (1H,dd,J=12,6Hz),3.97(2H,m),4.25(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 6.99 min

EXAMPLE 18 (1R,5S,6S)-2-[(2S,4S)-2-(2,5-dioxopyrrolidin-3-yl)pyrrolidin-4-ylthio]-6,[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid ##STR1160##

The same procedure as in Example 8-1 was carried out by using allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (230 mg, 0.46 mmol) and (2S,4S)-N-allyloxycarbonyl-2-(2,5-dioxopyrrolidin-3-yl)-4-mercaptopyrrolidine (130 mg, 0.46 mmol, compound of Reference Example 9) to obtain allyl (1R,5S,6S)-2-[(2S,4S)-N-allyloxycarbonyl-2-(2,5-dioxopyrrolidin-3-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (125 mg, yield: 50.9%).

IR(KBr)cm⁻¹ : 1780, 1720, 1410, 1330

NMR(CDCl₃) δ: 1.24(3H,d,J=7Hz),1.33(3H,d,J=6Hz),1.63(1H,m), 5.13-5.54(4H,m),5.78-6.08(2H,m) ##STR1161##

The same procedure as in Example 8-2 was carried out by using the compound obtained by the above reaction. (120 mg, 0.23 mmol ) to obtain the above identified compound (28.5 mg, yield: 30.9%).

IR(KBr)cm⁻¹ : 1760, 1720, 1600, 1380

NMR(D₂ O) δ: 1.18(3H,d,J=7Hz),1.25(3H,d,J=6Hz),1.70(1H,m), 2.53-2.80(2H,m),3.03(1H,m),3.25-3.50(4H,m), 3.60(1H,m),3.80-4.00(2H,m),4.20(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 2.46 min

EXAMPLE 19 (5R,6S),6,[(R)-1-hydroxyethyl]-2-[(2S,4S)-2-3-pyrazolidinon-5-yl )pyrrolidin-4-ylthio ]-1-carbapen-2-em-3-carboxylic acid ##STR1162##

The same procedure as in Example 8-1 was carried out by using allyl (5R,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (195 mg, 0.40 mmol) and (2S,4S)-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-3-pyrazolidinon-5-yl)-4-mercaptopyrrolidine (162 mg, 0.40 mmol) to obtain allyl (5R,6S)-2-[(2S,4S)-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-3-pyrazolidinon-5-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (155 mg, yield: 65.4%).

IR(KBr)cm⁻¹ : 1780, 1700, 1550, 1410, 1330

NMR(CDCl₃) δ: 1.32(3H,d,J=6Hz),4.52-4.90(6H,m),5.18-5.56 (6H,m),5.78-6.10(3H,m) ##STR1163##

The same procedure as in Example 8-2 was carried out by using the compound obtained by the above reaction (155 mg, 0.26 mmol) to obtain the above identified compound (39 mg, yield: 38.8%).

IR(KBr)cm⁻¹ : 1760, 1680, 1590, 1390

NMR(CDCl₃) δ: 1.26(3H,d,J=6Hz),1.88(1H,m),2.35(1H,m),2.70(1H,m), 3.06-3.27(2H,m),3.42(2H,m),3.70-3.90(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 1.53 min

EXAMPLE 20 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(3-pyrazolidinon-5-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid ##STR1164##

The same procedure as in Example 8-1 was carried by using allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (330 mg, 0.66 mmol) and (2S,4S)-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-3-pyrazolidinon-5-yl)-4-mercaptopyrrolidine (234 mg, 0.66 mmol) to obtain allyl (1R,5S,6S)-2-[(2S,4S)-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-3-pyrazolidinon-5-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (172 mg, yield: 43.1%).

IR(KBr)cm⁻¹ : 1770, 1700, 1410, 1320

NMR(CDCl₃) δ: 1.24(3H,d,J=7Hz),1.35(3H,d,J=6Hz),4.50-4.90 (6H,m),5.20-5.56(6H,m ),5.80-6.10 (3H,m) ##STR1165##

The same procedure as in Example 8-2 was carried out by using the compound obtained by the above reaction (172 mg, 0.285 mmol) to obtain the above identified compound (42 mg, yield: 37.2%).

IR(KBr)cm⁻¹ : 1760, 1680, 1590, 1390

NMR(D₂ O) δ: 1.20(3H,d,J=7Hz),1.26(3H,d,J=6Hz),1.85(1H,m),2.35 (1H,br d,J=18Hz),2.70(1H,m),3.10(1H,dd,J=18,9Hz), 3.15-3.52(3H,m),3.62-3.90(2H,m),3.93-4.32(4H,m)

HPLC (the same condition as in Example 1)

Retention time: 2.23 min

EXAMPLE 21 Sodium (5R,6S)-6-[(R)-1-hydroxyethyl]-2-[(2S,4S)-2-(2-pyrrolidon-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate ##STR1166##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (5R,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (180 mg, 0.31 mmol) and (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-3-yl)pyrrolidine (110 mg, 0.30 mmol, compound of Reference Example 11) to obtain p-nitrobenzyl (5R,6S)-6-[(R)-1-hydroxyethyl]-2-(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (151 mg, yield: 70%).

IR(KBr)cm⁻¹ : 1780, 1700, 1520, 1350

NMR(CDCl₃) δ: 1.38(3H,d,J=6Hz),5.30(3H,m),5.56(1H,d,J=14Hz),7.57 (2H,d,J=8Hz),7.70(2H,d,J=8Hz),8.25(2H,d,J=8Hz), 8.28(2H,d,J=8Hz) ##STR1167##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (150 mg, 0.22 mmol) to obtain the above identified compound (35 mg, yield: 38.1%).

IR(KBr)cm⁻¹ : 1760, 1690, 1590, 1380

NMR(D₂ O) δ: 1.29(3H,d,J=6Hz),1.72-2.07(3H,m),2.39(1H,m), 2.80(1H,m),3.00(1H,m),3.20(2H,m),3.44 (3H,m), 3.78(2H,m), 3.99(1H,m), 4.24(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 1.93 min

EXAMPLE 22 Sodium (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(2-pyrrolidon-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate ##STR1168##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (180 mg, 0.30 mmol) and (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon- 3-yl)pyrrolidine (110 mg, 0.30 mmol) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (161 mg, yield: 74.9%).

IR(KBr)cm⁻¹ : 1780, 1700, 1520, 1350

NMR(CDCl₃) δ: 1.32(3H,d,J=7Hz),1.40(3H,d,J=6Hz),5.32(3H,m),5.56 (1H,d,J=14Hz),7.58(2H,d,J=8Hz),7.70(2H,d,J=8Hz), 8.27(2H,d,J=8Hz),8.29(2H,d,J=8Hz) ##STR1169##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (160 mg, 0.23 mmol) to obtain the above identified compound (37 mg, yield: 37.5%).

IR(KBr)cm⁻¹ : 1760, 1690, 1600, 1380

NMR(D₂ O) δ: 1.18(3H,d,J=7Hz), 1.25(3H,d,J=6Hz), 1.63-2.07 (3H,m), 2.36(1H,m),2.74(1H,m),2.98(1H,m), 3.30-3.50(4H,m), 3.72(2H,m), 3.95(1H,m), 4.22(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 3.23 min

EXAMPLE 23 (1R,5S,6S)-2-[(2S,4S)-2-(2-carbamoylpyrrolidin-4-yl)pyrrolidin-4-ylthio ]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid diastereomer I ##STR1170##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (174 mg, 0.29 mmol) and (2S,4S)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer I (160 mg, 0.28 mmol, compound of Reference Example 13) to obtain p-nitrobenzyl (1R,5S,6S)-2-[(2S,4S)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer I (223 mg, yield: 82.9%).

IR(KBr)cm⁻¹ : 1770, 1700, 1520, 1400, 1350

NMR(CDCl₃) δ: 1.28(3H,d,J=7Hz),1.37(3H,d,J=6Hz),5.25(5H,m),5.52 (1H,d,J=14H),7.53(4H,d,J=8Hz),7.67 (2H,d,J=8Hz), 8.23(6H,d,J=8Hz) ##STR1171##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (223 mg, 0.24 mmol) to obtain the above identified compound (50.7 mg, yield: 49.8%).

IR(KBr)cm⁻¹ : 1750, 1700, 1390

NMR(D₂ O) δ: 1.19(3H,d,J=7Hz),1.27(3H,d,J=6Hz),1.58(2H,m), 2.00-2.32(2H,m),2.42-2.74(2H,m),2.82 (1H,dd,J=12,8Hz),3.22(1H,dd,J=12,4Hz), 3.30-3.54(4H,m),3.92(1H,m),4.10(1H,dd,J=9,4Hz), 4.20(2H,m)

EXAMPLE 24 (1R,5S,6S)-2-[(2S,4S)-2-(2-carbamoylpyrrolidin-4-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid diastereomer II ##STR1172##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (290 mg, 0.49 mmol) and (2S,4S)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer II (280 mg, 0.49 mmol, compound of Reference Example 14) to obtain p-nitrobenzyl (1R,5S,6S)-2-[(2S,4S)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer II (419 mg, yield: 93.4%).

IR(KBr)cm⁻¹ : 1770, 1700, 1520, 1350

NMR(CDCl₃) δ: 1.28(3H,d,J=7Hz),1.37(3H,d,J=6Hz),5.26(5H,m),5.52 (1H,d,J=14Hz),7.53(4H,d,J=8Hz),7.68(2H,d,J=8Hz), 8.24(6H,d,J=8Hz) ##STR1173##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (419 mg, 0.46 mmol) to obtain the above identified compound (93 mg, yield: 48%).

IR(KBr)cm⁻¹ : 1760, 1700, 1390

NMR(D₂ O) δ: 1.20(3H,d,J=7Hz),1.28(3H,d,J=6Hz),1.62(2H,m), 2.44-2.75(3H,m),2.83(1H,dd,J=10,8Hz), 3.16-3.57(6H,m),3.94(1H,m),4.08(1H,m),4.23(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 2.84 min

EXAMPLE 25 (5R,6S)-6-[(R)-1-hydroxyethyl]-2-[(2S,4S)-2-(pyrrolidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid diastereomer A ##STR1174##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (5R,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (255 mg, 0.45 mmol) and (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidine (240 mg, 0.45 mmol, compound of Reference Example 5) to obtain p-nitrobenzyl (5R,6S)-6-[(R)-1-hydroxyethyl]-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidin-4ylthio]-1-carbapen-2-em-3-carboxylate (297 mg, yield: 78.6%).

IR(KBr)cm⁻¹ : 1780, 1700, 1520, 1350

NMR(CDCl₃) δ: 1.25(3H,d,J=6Hz),4.00-4.40(4H,m),5.16-5.35(5H,m), 5.52(1H,d,J=14Hz),7.52(4H,d,J=8Hz),7.65(2H,d,J=8Hz), 8.21(6H,m) ##STR1175##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (297 mg, 0,345 mmol) to obtain diastereomer A (13.6 mg, yield: 10.7%) of the above identified compound.

IR(KBr)cm⁻¹ : 1760, 1690, 1390

NMR(D₂ O) δ: 1.27(3H,d,J=6Hz),1.46(1H,m),1.68-2.10(2H,m), 2.14-2.80(3H,m),3.80(2H,m),4.22(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 2.03 min

EXAMPLE 26 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2[(2S,4S)-2-(pyrrolidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid diastereomer A ##STR1176##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (1.75 g, 2.94 mmol) and (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidine diastereomer A (1.56 g, 2.94 mmol, compound of Reference Example 15-7) to obtain p-nitrobenzyl (1R,5S,6S]i-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)- 2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate diastereomer A (2.37 g, yield: 92%).

IR(KBr)cm⁻¹ : 1780, 1700, 1520, 1350

NMR(CDCl₃) δ: 1.28(3H,t,J=7Hz),1.37(3H,d,J=6Hz),1.60-2.15(4H,m), 2.55(1H,m),2.78(1H,m),4.04-4.35(3H,m),5.24(5H,m), 5.53(1H,br d,J=14Hz),7.54(4H,br d,J=8Hz), 7.67(2H,d,J=8Hz),8.24(6H,br d,J=8Hz) ##STR1177##

The same procedure as in Example 11-2 was carried out by using the compound obtained by the above reaction (2.37 g, 2.71 mmol) to obtain the above identified compound (298 mg, yield: 28.8%). Various spectra data of this compound agreed to the data of the diastereomer A obtained in Example 11-2.

EXAMPLE 27 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2[(2S,4S)-2-(pyrrolidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid diastereomer B ##STR1178##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (1.27 g, 2.14 mmol) and (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidine diastereomer B (1.14 g, 2.15 mmol, compound of Reference Example 16) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidin-4ylthio]-1-carbapen-2-em-3-carboxylate diastereomer B (1.65 g, yield: 88.3%).

IR(KBr)cm⁻¹ : 1780, 1700, 1520, 1350

NMR(CDCl₃) δ: 1.28(3H,t,J=7Hz),1.37(3H,t,J=6Hz),1.50-2.08(4H,m), 2.42-2.70(2H,m),3.48-3.70(3H,m),4.01-4.76(3H,m), 5.24(5H,m),5.54(1H,d,J=14Hz),7.54(4H,br d,J=8Hz), 7.67(2H,d,J=8Hz),8.24(6H,br d,J=8Hz) ##STR1179##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (1.65 g, 1.89 mmol) to obtain the above identified compound (220 mg, yield: 30.6%). Various spectra data of this compound agreed to those of the diastereomer B obtained in Example 11-2.

EXAMPLE 28 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2S,4S)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid diastereomer A ##STR1180##

The same procedure as in Example 1-1 was carried cut by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen- 2-em-3-carboxylate (6.67 g, 11.2 mmol) and (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer A (3.9 g, 10.7 mmol, compound of Reference Example 18-4) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate diastereomer A (6.01 g, yield: 77.1%).

IR(KBr)cm⁻¹ : 1770, 1700, 1520, 1340, 1250

NMR(CDCl₃) δ: 1.28(3H,d,J=8Hz),1.33(3H,d,J=7Hz),1.72(2H,m), 2.06-2.68(3H,m),2.96-3.7(5H,m),4.04-4.36(2H,m), 5.24(2H,m),5.31 and 5.52(2H,ABq,J=14Hz),5.8(1H,br), 7.53(2H,d,J=8Hz),7.64(2H,d,J=8Hz),8.20(2H,d,J=8Hz), 8.22(2H,d,J=8Hz) ##STR1181##

The same procedure as in Example 1-2 was carried out by using compound obtained by the above reaction (6 g, 8.62 mmol) to obtain the above identified compound (1.68 g, yield: 49.4%).

IR(KBr )cm⁻¹ : 1755, 1680, 1595, 1385, 1280

NMR(D₂ O) δ: 1.15(3H,d,J=8Hz),1.23(3H,d,J=7Hz),1.7(1H,m), 2.3(1H,m),2.54-3.1(3H,m),3.1-3.5(3H,m), 3.55-3.94(3H,m),4.0(1H,m),4.19(2H,m)

HPLC:

Column: YMC AQ-304

Eluent: 0.01M Phosphate buffer (pH 7.0)-acetonitrile (96:4)

Flow rate: 1.0 ml/min

Temperature: 40° C.

Detector: 254 nm

Retention time: 14.83 min

EXAMPLE 29 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid diastereomer B ##STR1182##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen- 2-em-3-carboxylate (10.61 g, 17 mmol) and (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer B (5.92 g, 16.2 mmol, compound of Reference Example 19-4) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate diastereomer B (9.9 g, yield: 83.7%).

IR(KBr)cm⁻¹ : 1770, 1705, 1695, 1605, 1345, 1205

NMR(D₂ O) δ: 1.27(3H,d,J=8Hz),1.36(3H,d,J=7Hz),1.72-2.06(2H,m), 2.1-2.68(3H,m),3.02-3.7(5H,m),4.06-4.16(2H,m), 5.31 and 5.52(2H,ABq,J=14Hz),6.04(1H,br),7.53 (2H,d,J=8Hz),7.66(2H,d,J=8Hz),8.20(2H,d,J=8Hz), 8.22(2H,d,J=8Hz) ##STR1183##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (9.8 g, 14.1 mmol) to obtain the above identified compound (1.42 g, yield: 25.5%).

IR(KBr)cm⁻¹ : 1755, 1680, 1590, 1390, 1280

NMR(D₂ O) δ: 1.19(3H,d,J=8Hz),1.26(3H,d,J=7Hz),1.72(1H,m), 2.28(1H,m) ,2.52-3.10(3H,m),3.26-3.50(3H,m), 3.60-3.90(3H,m),4.02(1H,m),4.22(2H,m)

HPLC:

Column: YMC AQ-304

Eluent: 0.01M Phosphate buffer (pH 7.0)-acetonitrile (96:4)

Flow rate: 1.0 ml/min

Temperature: 40° C.

Detector: 254 nm

Retention time: 15.18 min

EXAMPLE 30 (1R,5S,6S)-2-[(2S,4S)-2-(3-amino-2-pyrrolidon-4-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl-1-methyl-1-carbapen-2-em-3-carboxylic acid ##STR1184##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen- 2-em-3-carboxylate (155.4 mg, 0.26 mmol) and (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[3-[(p-nitrobenzyloxycarbonyl)amino]-2-pyrrolidon-4-yl]pyrrolidine (110 mg, 0.197 mmol, compound of Reference Example 20-6) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[3-[(p-nitrobenzyloxycarbonyl)amino]-2-pyrrolidon-4-yl]pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (87 mg, yield: 48.9%).

IR(KBr)cm⁻¹ : 1775, 1710, 1605, 1525, 1350, 735

NMR(CDCl₃) δ: 1.27(3H,d,J=8Hz),1.35(3H,d,J=8Hz),1.6-1.8(2H,m), 2.78(1H,m),3.1-3.5(6H,m),3.65(1H,m),4.0-4.4(5H,m), 5.2(4H,s),5.28(1H,d,J=16Hz),5.5(1H,d,J=16Hz),6.32 (1H,br),7.52(4H,d,J=9Hz),7.65(2H,d,J=8Hz),8.2(6H,m) ##STR1185##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (87 mg, 0.096 mmol). The obtained reaction solution was subjected to column chromatography (HP-20 SS, 5-10% methanol aqueous solution), and the desired fraction was concentrated and freeze-dried to obtain the above identified compound (25.72 mg, yield: 65.1% ).

IR(KBr)cm⁻¹ : 1755, 1700, 1605, 1595, 1385

NMR(D₂ O) δ: 1.17(3H,d,J=8Hz),1.24(3H,d,J=8Hz),2.5-2.8(2H,m), 2.6-2.8(2H,m),3.1-3.6(5H,m),3.9-4.4(5H,m)

EXAMPLE 31 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2[(2R,4S)-2-(pyrrolidin-3-ylmethyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid ##STR1186##

The same procedure as in Example 8-1 was carried out by using allyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-diphenoxyphosphoryloxy-1-carbapen-2-em-3-carboxylate (790 mg, 1.6 mmol), (2R,4S)-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-3-ylmethyl)-4-mercaptopyrrolidine (560 mg, 1.6 mmol) and N,N-diisopropylethylamine (0.28 ml, 1.6 mmol) to obtain allyl (1R,5S,6S)-2-[(2R,4S)-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-3-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (480 mg, yield: 50%).

NMR(CDCl₃) δ: 1.28(3H,d,J=6Hz),1.36(3H,d,J=6Hz),1.5-1.8(3H,m), 1.9-2.2(4H,m),2.62(1H,m),3.02(1H,m),3.2-3.4(4H,m), 3.5-3.7(3H,m),4.0-4.3(3H,m),4.6-4.9(6H,m), 5.2-5.56(6H,m),5.9-6.1(3H,m) ##STR1187##

The same procedure as in Example 8-2 was carried out by using the compound obtained by the above reaction (480 mg, 0.8 mmol), bis(triphenylphosphine)palladium(II) chloride (11 mg, 0.016 mmol), tributyltin hydride (1.28 ml, 4.77 mmol) and water (107 μl, 5.96 mmol). The obtained aqueous layer was purified by reversed phase column chromatography (YMC-GEL™ ODS-AQ 120-S50, 50 ml, eluted with methanol-water (1:4)) and freeze-dried to obtain the above identified compound (128 mg, yield: 41%).

IR(KBr)cm⁻¹ : 1760, 1600, 1390

NMR(D₂ O) δ: 1.2(3H,d,J=6Hz),1.28(3H,d,J=7Hz),1.6-1.9(3H,m), 2.2-2.7(3H,m),2.9-3.6(10H,m),3.8(1H,m),4.2-4.4(2H,m)

EXAMPLE 32 (1R, 5S,6S)-2-[(2R,4S)-2-[(2S)-2-carbamoylpyrrolidin-4-ylmethyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid ##STR1188##

The same procedure as in Example 8-1 was carried out by using allyl (IR,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-diphenoxyphosphoryloxy-1-carbapen-2-em-3-carboxylate (264 mg, 0.49 mmol), (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-carbamoylpyrrolidin-4-ylmethyl]-4-mercaptopyrrolidine (210 mg, 0.49 mmol) and N,N-diisopropylethylamine (92 μl, 0.49 mmol) to obtain allyl (1R,5S,6S)-2-[(2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-carbamoylpyrrolidin-4-ylmethyl]pyrrolidin-4-ylthio]-6-(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (250 mg, yield: 73%).

NMR(CDCl₃) δ: 1.25(3H,d,J=6Hz),1.34(3H,d,J=7Hz),1.5-1.8(3H,m), 1.8-2.7(4H,m),3.0-3.4(4H,m),3.5-4.4(7H,m), 4.5-4.9(6H,m),5.2-5.5(7H,m),5.8-6.1(3H,m),6.9 (0.5H,br s),7.3(0.5H,br s) ##STR1189##

The same procedure as in Example 8-2 was carried out by using the compound obtained by the above reaction (250 mg, 0.387 mmol), bis(triphenylphosphine)palladium(II) chloride (5.4 mg, 0.008 mmol), tributyltin hydride (0.62 ml, 2.3 mmol) and water (52 μl, 2.9 mmol). The obtained aqueous layer was purified by reversed phase column chromatography (LC-SORB™ SP-B-ODS, 14 ml, eluted with methanol-water (3:7)) and freeze-dried to obtain the above identified compound (107 mg, yield: 63%).

IR(KBr)cm⁻¹ : 1750, 1680, 1590, 1390

NMR(D₂ O) δ: 1.22(3H,d,J=7Hz),1.28(3H,d,J=6Hz),1.58(1H,m), 1.8-2.4(5H,m),2.6-2.84(2H,m),3.2-3.7(6H,m), 3.95(1H,m),4.1(1H,dd,J=10,4Hz),4.2-4.3(2H,m)

EXAMPLE 33 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2R,4S)-2-[(2S)-2-(methylcarbamoyl)pyrrolidin-4-ylmethyl]pyrrolidin-4-ylthio]-]-carbapen-2-em-3-carboxylic acid ##STR1190##

The same procedure as in Example 8-1 was carried out by using allyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-diphenoxyphosphoryloxy-1-carbapen-2-em-3-carboxylate (204 mg, 0.41 mmol), (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-(methylcarbamoyl)pyrrolidin-4-ylmethyl]-4-mercaptopyrrolidine (168 mg, 0.41 mmol) and N,N-diisopropylethylamine (71 μl, 0.41 mmol) to obtain allyl (1R,5S,6S)-2-[(2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-(methylcarbamoyl)pyrrolidin-4-ylmethyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (180 mg, yield:

NMR(CDCl₃) δ: 1.26(3H,d,J=6Hz),1.36(3H,d,J=6Hz),1.5-1.8(3H,m), 1.8-2.6(4H,m),2.8(3H,d,J=4Hz),3.0(1H,m), 3.1-3.4(3H,m),3.8(2H,m),3.9-4.4(5H,m), 4.5-4.9(6H,m),5.1-5.5(6H,m),5.8-6.0(3H,m), 6.9(0.5H,br s),7.3(0.5H,br s) ##STR1191##

The same procedure as in Example 8-2 was carried out by using the compound obtained by the above reaction (180 mg, 0.27 mmol), bis(triphenylphosphine)palladium(II) chloride (4 mg, 0.005 mmol), tributyltin hydride (0.44 ml, 1.6 mmol) and water (37 μl, 2 mmol). The obtained aqueous layer was purified by reversed phase column chromatography (LC-SORB™ SP-B-ODS, 14 ml, eluted with methanol-water (3:7)) and feeze-dried to obtain the above identified compound (60 mg, yield: 49%).

IR(KBr)cm⁻¹ : 1750, 1600, 1390

NMR(D₂ O) δ: 1.2(3H,d,J=6Hz),1.28(3H,d,J=7Hz),1.56(1H,m), 1.8-2.3(5H,m),2.6-2.8(2H,m),2.76(3H,s), 3.2-3.7(6H,m),3.95(1H,m),4.04(1H,dd,J=10,4Hz), 4.2-4.3(2H,m)

EXAMPLE 34 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2R,4S)-2-[(2S)-2-(dimethylcarbamoyl)pyrrolidin-4-ylmethyl]pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid ##STR1192##

The same procedure as in Example 8-1 was carried out by using allyl (IR,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-diphenoxyphosphoryloxy-1-carbapen-2-em-3-carboxylate (129 mg, 0.26 mmol), (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-(dimethylcarbamoyl)pyrrolidin-4-ylmethyl]-4-mercaptopyrrolidine (110 mg, 0.26 mmol) and N,N-diisopropylethylamine (45 μl, 0.26 mmol) to obtain allyl (1R,5S,6S)-2-[(2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-(dimethylcarbamoyl)pyrrolidin-4-ylmethyl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (78 mg, yield: 45%).

NMR(CDCl₃) δ: 1.26(3H,d,J=6Hz),1.37(3H,d,J=6Hz),1.4-2.6(7H,m), 2.9-3.3(4H,m),2.98(3H,s),3.12(3H,s),3.5-4.3 (7H,m),4.5-4.9(6H,m),5.1-5.5(6H,m),5.9(3H,m) ##STR1193##

The same procedure as in Example 8-2 was carried out by using the compound obtained by the above reaction (78 mg, 0.12 mmol), bis(triphenylphosphine)palladium(II) chloride (2 mg, 0.002 mmol), tributyltin hydride (0.186 ml, 0.69 mmol) and water (16 μl, 0.87 mmol). The obtained aqueous layer was purified by reversed phase column chromatography (LC-SORB™ SP-B-ODS, 14 ml, eluted with methanol-water (2:3)) and freeze-dried to obtain the above identified compound (11 mg, yield: 20%).

IR(KBr)cm⁻¹ : 1750, 1640, 1600, 1390

NMR(D₂ O) δ: 1.2(3H,d,J=6Hz),1.28(3H,d,J=7Hz),1.5(1H,m),1.8-2.4 (5H,m),2.6-2.9(2H,m),2.96(3H,s),3.06(3H,s),3.1-3.6 (6H,m),3.8-4.0(2H,m),4.1-4.3(2H,m)

EXAMPLE 35 (1R,5S,6S)-2-[(2S,4S)-2-(2,4-dioxoimidazolidin-5-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl)]-1-methyl-1-carbapen-2-em-3-carboxylic acid ##STR1194##

(2S,4S)-2-(2,4-dioxoimidazolidin-5-yl)-4-(p-methoxybenzylthio)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (250 mg, 0.499 mmol, compound of Reference Example 28) and anisole (0.27 ml, 2.48 mmol ) were dissolved in trifluoroacetic acid (5 ml), and the solution was cooled with ice under a nitrogen stream. Then, trifluoromethanesulfonic acid (0.07 ml, 0.791 mmol) was dropwise added thereto, and the mixture was stirred for 50 minutes under cooling with ice. The solvent was distilled off, and the residue was washed with diethyl ether to obtain a crude thiol.

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (300 mg, 0.505 mmol) and the thiol obtained by the above reaction to obtain p-nitrobenzyl (1R,5S,6S)-2-[(2S,4S)-2-(2,4-dioxoimidazolidin-5-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-6-[(R)-1hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (100 mg, yield: 28%).

IR(KBr)cm⁻¹ : 3430, 1775, 1725, 1610, 1520, 1350, 1285, 1250

NMR(CDCl₃) δ: 1.27(3H,d,J=6Hz),1.33(3H,d,J=6Hz),4.4(1H,m), 4.7 and 5.05(1H,s),5.25(2H,s),5.3 and 5.5 (2H,ABq,J=14Hz),7.58(2H,d,J=8Hz),7.69(2H,d,J=8Hz), 8.2-8.3(4H,m) ##STR1195##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (100 mg, 0.138 mmol) to obtain the above identified compound (23 mg, yield: 41%).

IR(KBr)cm⁻¹ : 3440, 1760, 1730, 1600, 1400

NMR(D₂ O) δ: 1.20(3H,d,J=7Hz),1.27(3H,d,J=7Hz),1.75(1H,m), 2.6(1H,m),3.0-3.6(4H,m),3.7-4.0(2H,m),4.1-4.3 (2H,m),4.57(1H,d,J=8Hz)

UV λ_(max) (0.1M 3-morpholinopropanesulfonic acid buffer pH 7.0):300 nm (ε=9400)

EXAMPLE 36 (5R,6S)-2-[(2S,4S)-2-(2,4-dioxoimidazolidin-5-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylic acid ##STR1196##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (5R,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (150 mg, 0.258 mmol) and (2S,4S)-2-(2,4-dioxoimidazolidin-5-yl)-4-(p-methoxybenzylthio)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (120 mg, 0.240 mmol, compound of Reference Example 28) to obtain p-nitrobenzyl (5R,6S)-2-[(2S,4S)-2-(2,4-dioxoimidazolidin-5-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (30 mg, yield: 18%).

IR(KBr)cm⁻¹ : 3400, 1780, 1730, 1610, 1520, 1350

NMR(CDCl₃ +CD₃ OD) δ: 1.32(3H,d,J=6Hz),1.78(1H,m),2.40(1H,m),3.0-3.5(4H,m), 4.0-4.5(4H,m),4.73 and 5.04(1H,s),5.28(2H,s), 5.28 and 5.49(2H,ABq,J=13Hz),7.57(2H,d,J=9Hz),7.67 (2H,d,J=9Hz),8.1-8.3(4H,m) ##STR1197##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (30 mg, 0.042 mmol) to obtain the above identified compound (11 mg, yield: 66%).

IR(KBr)cm⁻¹ : 3430, 1760, 1720, 1600, 1400

NMR(D₂ O) δ: 1.26(3H,d,J=7Hz),1.55(1H,m),2.5(1H,m),3.1-3.6(4H,m), 4.1-4.3(2H,m),4.38(1H,d,J=5Hz)

UV λ_(max) (0.1M 3-morpholinopropanesulfonic acid buffer pH 7.0):301 nm (ε=4600)

EXAMPLE 37 (1R,5S,6S)-2-[(2R,4S)-2-(2,4-dioxoimidazolidin-5-ylmethyl )pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid diastereomer A ##STR1198##

The same procedure as in Example 8-1 was carried out by using allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (220 mg, 0.440 mmol) and (2R,4S)-N-allyloxycarbonyl-2-(2,4-dioxoimidazolidin-5-ylmethyl)-4-tritylthiopyrrolidine diastereomer A (270 mg, 0.498 mmol, compound of Reference Example 29) to obtain allyl (1R,5S,6S)-2-[(2R,4S)-N-allyloxycarbonyl-2-(2,4-dioxoimidazolidin-5-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer A (70 mg, yield: 26%).

IR(KBr)cm⁻¹ : 3550, 3480, 3420, 3200, 1780, 1730, 1550,

NMR(CDCl₃) δ: 1.26(3H,d,J=7Hz),1.36(3H,d,J=7Hz),1.5-2.5(4H,m), 2.7(2H,m),3.1-3.6(3H,m),3.7(1H,m),3.8-4.4(4H,m), 4.4-4.9(4H,m),5.0-5.5(4H,m),5.6-6.05(2H,m), 6.89(1H,s),8.78(1H,s) ##STR1199##

The same procedure as in Example 8-2 was carried out by using the compound obtained by the above reaction (70 mg, 0.128 mmol) to obtain the above identified compound 19 mg, yield: 35%).

IR(KBr)cm⁻¹ : 3400, 3250, 1760, 1720, 1590, 1390

NMR(D₂ O) δ: 1.15(3H,d,J=6Hz),1.22(3H,d,J=7Hz),1.7(1H,m),2.3 (2H,m),2.75(1H,m),3.2-3.5(3H,m),3.5-3.9(2H,m), 3.95(1H,m),4.2(2H,m),4.34(1H,t,J=6Hz)

UV λ_(max) (0.1M 3-morpholinopropanesulfonic acid buffer pH 7.0):298 nm (ε=9300)

EXAMPLE 38 (1R,5S,6S)-2-[(2R,4S)-2-(2,4-dioxoimidazolidin-5-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid diastereomer ##STR1200##

The same procedure as in Example 8-1 was carried out by using allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (220 mg, 0.440 mmol) and (2R,4S)-N-allyloxycarbonyl-2-(2,4-dioxoimidazolidin-5-ylmethyl)-4-tritylthiopyrrolidine diastereomer B (220 mg, 0.406 mmol, compound of Reference Example 29) to obtain allyl (1R,5S,6S)-2-[(2R,4S)-N-allyloxycarbonyl-2-(2,4-dioxoimidazolidin-5-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer B (70 mg, yield: 31%).

IR(KBr)cm⁻¹ : 3420, 3270, 1775, 1730, 1550, 1415

NMR(CDCl₃) δ: 1.27(3H,d,J=7Hz),1.36(3H,d,J=7Hz),1.5-2.1(4H,m), 2.3(1H,m),2.7(1H,m),3.1-3.45(3H,m),3.65(1H,m), 3.9-4.4(4H,m),4.4-4.9(4H,m),5.1-5.6(4H,m),5.7-6.1 (2H,m),6.73(1H,s),8.34(1H,s) ##STR1201##

The same procedure as in Example 8-2 was carried out by using the compound obtained by the above reaction (70 mg, 0.128 mmol) to obtain the above identified compound (18 mg, yield: 33%).

IR(KBr)cm⁻¹ : 3420, 3230, 1760, 1720, 1590, 1395

NMR(D₂ O) δ: 1.12(3H,d,J=7Hz),1.19(3H,d,J=7Hz),1.65(1H,m), 2.3(2H,m),2.73(1H,m),3.2-3.5(3H,m),3.61(1H,m), 3.8(1H,m),3.94(1H,m),4.15(2H,m),4.34(1H,t,J=6Hz)

UV λ_(max) (0.1M 3-morpholinopropanesulfonic acid buffer pH 7.0):298 nm (ε=9400)

EXAMPLE 39 Sodium (1R,5S,6S)-2-[(2R,4S)-2-(2,5-dioxopyrrolidin-3-ylmethyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate ##STR1202##

A 2N sodium hydroxide aqueous solution (0.34 ml, 0.68 mmol) was added to a solution of (2R,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-(2,5-dioxopyrrolidin-3-ylmethyl)pyrrolidine (250 mg, 0.57 mmol, compound of Reference Example 30) in methanol (5.0 ml) under cooling with ice, and the mixture was stirred for 30 minutes. Then, 6N hydrochloric acid (0.11 ml, 0.66 mmol) was added thereto, and the mixture was extracted with methylene chloride. The extract was dried over anhydrous magnesium sulfate and concentrated to obtain (2R,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2,5-dioxopyrrolidin-3ylmethyl)pyrrolidine.

The same procedure as in Example 1-1 was carried out by using the thiol obtained by the above reaction and p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (273 mg, 0.46 mmol) to obtain p-nitrobenzyl (1R,5S,6S)-2-[(2R,4S)-2-(2,5-dioxopyrrolidin-3-ylmethyl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (229 mg, yield: 67%).

NMR(CDCl₃) δ: 1.28(3H,d,J=8Hz),1.38(3H,d,J=8Hz),5.24(4H,br s), 7.54(4H,d,J=8Hz),8.25(4H,d,J=8Hz) ##STR1203##

The same procedure as in Example 1-2 was conducted by using the compound obtained by the above reaction (210 mg, 0.28 mmol) to obtain the above identified compound (26 mg, yield: 21%).

IR(KBr)cm⁻¹ : 3400, 2950, 1760, 1720, 1600, 1380, 1180

NMR(D₂ O) δ: 1.10 (3H,d,J=6Hz),1.16(3H, d,J=6Hz),1.58 (1H,m),2.00 (1H,m),2.46(1H,m),2.66(1H,m),2.94(2H,m),3.28(2H,m), 3.54(1H,m),3.74(1H,m ),4.12(2H,m)

HPLC:

Eluent: 0.01M Phosphate buffer (pH 6.5)-methanol (85:15)

Flow rate: 0.8 ml/min

Other conditions are the same as in Example 1

Retention time: 4.28 min

EXAMPLE 40 (1R,5S,6S)-2-[(2S,4S)-2-(5-oxopiperazin-2-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid diastereomer A ##STR1204##

The same procedure as in Example 39-1 was carried out by using (2S,4S)-N-allyloxycarbonyl-4-acetylthio-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)pyrrolidine diastereomer A (250 mg, 0.60 mmol, compound of Reference Example 31) to obtain (2S,4S)-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)-4-mercaptopyrrolidine diastereomer A.

The same procedure as in Example 1-1 was carried out by using the thiol obtained by the above reaction and allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]- 1-methyl-1-carbapen-2-em-3-carboxylate (242 mg, 0.48 mmol) to obtain allyl (1R,5S,6S)-2-[(2S,4S)-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer A (222 mg, yield: 74%).

NMR(CDCl₃) δ: 1.26(3H,d,J=6Hz),1.38(3H,d,J=6Hz),1.80(1H,m), 2.50(1H,m),3.0-5.0(19H,m),5.20-5.60(6H,m),5.96(3H,m) ##STR1205##

The same procedure as in Example 9-2 was conducted by using the compound obtained by the above reaction (diastereomer A, 220 mg, 0.35 mmol) to obtain the above identified compound (diastereomer A, 88 mg, yield: 60%).

IR(KBr)cm⁻¹ : 3400, 2950, 1760, 1660, 1600, 1380

NMR(D₂ O) δ: 1.20(3H,d,J=6Hz),1.26(3H,d,J=6Hz),1.78(1H,m), 2.64(1H,m),3.10-3.80(10H,m),4.00(1H,m),4.22(2H,m)

HPLC:

Flow rate: 0.8 ml/min

Other conditions are the same as in Example 1 PG,173

Retention time: 3.02 min

EXAMPLE 41 (1R,5S,6S)-2-[(2S,4S)-2-(5-oxopiperazin-2-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid diastereomer B ##STR1206##

The same procedure as in Example 39-1 was carried out by using (2S,4S)-4-acetylthio-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)pyrrolidine diastereomer B (200 mg, 0.48 mmol, compound of Reference Example 31) to obtain (2S,4S)-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)-4-mercaptopyrrolidine diastereomer B.

The same procedure as in Example 1-1 was carried out by using the thiol obtained by the above reaction and allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (243 mg, 0.48 mmol) to obtain allyl (1R,5S,6S)-2-[(2S,4S)-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer B (246 mg, yield: 65% ).

NMR(CDCl₃) δ: 1.24(3H,d,J=6Hz),1.38(3H,d,J=6Hz),1.80(1H,m), 2.60(1H,m),3.10-3.80(10H,m),4.0-5.0(9H,m), 5.20-5.60(6H,m),5.96(3H,m) ##STR1207##

The same procedure as in Example 9-2 was carried out by using the compound obtained by the above reaction (diastereomer B, 240 mg, 0.39 mmol) to obtain the above identified compound (diastereomer B, 53 mg, yield: 33%).

IR(KBr)cm⁻¹ : 3400, 1760, 1660, 1600, 1380

NMR(D₂ O) δ: 1.18(3H,d,J=6Hz),1.20(3H,d,J=6Hz),1.76(1H,m), 2.64(1H,m),3.10-3.80(10H,m),3.94(1H,m),4.18(2H,m)

HPLC:

Eluent: 0.1M Phosphate buffer (pH 6.5)-methanol (85:15)

Flow rate: 0.8 ml/min

Other conditions are the same as in Example 1

Retention time: 3.92 min

EXAMPLE 42 (5R,6S)-6-[(R)-1-hydroxyethyl]-2-[(2S,4S)-2-(piperidin-4-)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid ##STR1208##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (5R,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-carbapen-2-em-3-carboxylate (270 mg, 0.46 mmol) and (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidin-4-yl]pyrrolidine (216 mg, 0.40 mmol, compound of Reference Example 32) to obtain p-nitrobenzyl (5R,6S)-6-[(R)-1-hydroxyethyl]-2[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidin-4-yl]pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (267 mg, yield: 65.6%).

IR(KBr)cm⁻¹ : 1780, 1700, 1520, 1340

NMR(CDCl₃) δ: 1.35(3H,d,J=6Hz),2.76(2H,m),3.54(1H,m),4.00(1H,m), 5.24(5H,m),5.52(1H,d,J=14Hz),7.52(4H,d,J=8Hz),7.66 (2H,d,J=8Hz),8.22(6H,m) ##STR1209##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (265 mg, 0.30 mmol ) to obtain the above identified compound (33 mg, yield: 28.5%).

IR(KBr)cm⁻¹ : 1760, 1590, 1390

NMR(D₂ O) δ: 1.26(3H,d,J=6Hz),1.31-1.56(3H,m),1.69(1H,m), 1.85-2.14(2H,m),2.50(1H,m),2.81-3.06(4H,m), 3.10(3H,m),3.40(3H,m),3.74(1H,m),4.20(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 3.06 min

EXAMPLE 43 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(piperidin-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid ##STR1210##

The same procedure as in Example 1-1 was carried by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (245 mg, 0.41 mmol) and (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidin-4-yl]pyrrolidine (216 mg, 0.40 mmol, compound of Reference Example 32) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidin-4-yl]pyrrolidin-4ylthio]-1-carbapen-2-em-3-carboxylate (279 mg, yield: 76.2%).

IR(KBr)cm⁻¹ : 1770, 1700, 1520, 1340

NMR(CDCl₃) δ: 1.28(3H,t,J=7Hz),1.36(3H,t,J=6Hz),1.44-1.84(3H,m), 5.15(5H,m),5.53(1H,d,J=14Hz),7.56(4H,d,J=8Hz), 7.68(2H,d,J=8Hz),8.24(6H,m) ##STR1211##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (275 mg, 0.31 mmol) to obtain the above identified compound (57 mg, yield: 46.6%).

IR(KBr)cm⁻¹ : 1750, 1590, 1390

NMR(D₂ O) δ: 1.19(3H,d,J=7Hz),1.27(3H,d,J=6Hz),1.28-1.58(3H,m), 1.70(1H,m),1.85-2.15(2H,m),2.46(1H,m),2.78-3.18 (5H,m),3.40(4H,m),3.76(1H,m),4.22(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 3.28 min

EXAMPLE 44 (1R,5S,6S)-2-[(2S,4S)-2-(2-carbamoylpyrrolidin-4-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid diastereomer III ##STR1212##

The same procedure as in Example 1-1 was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (185 mg, 0.31 mmol) and (2S,4S)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer III (175 mg, 0.31 mmol, compound of Reference Example 33) to obtain p-nitrobenzyl (1R,5S,6S)-2-[(2S,4S)-2-[ 2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (218 mg, yield: 76.3%).

IR(KBr)cm⁻¹ : 1770, 1700, 1600, 1520, 1340

NMR(CDCl₃) δ: 1.27(3H,d,J=7Hz),1.36(3H,d,J=6Hz),4.02-4.40(5H,m), 5.22(5H,m),5.52(1H,d,J=14Hz),7.50(4H,d,J=8Hz),7.66 (2H,d,J=8Hz),8.22(6H,m) ##STR1213##

The same procedure as in Example 1-2 was carried out by using the compound obtained by the above reaction (218 mg, 0.24 mmol) to obtain the above identified compound (46 mg, yield: 45.6%).

IR(KBr)cm⁻¹ : 1760, 1680, 1600, 1390

NMR(D₂ O) δ: 1.19(3H,d,J=7Hz),1.27 (3H,d,J=6Hz),1.40-1.68(2H,m), 2.45(2H,m),2.60(1H,m),2.82(1H,m),3.04-3.47(6H,m), 3.85(1H,m),3.98(1H,t,J=8Hz),4.21(2H,m)

HPLC (the same condition as in Example 1)

Retention time: 2.15 min

EXAMPLE 45 (1R, 5S, 6S)-2 -[(2S,48)-2-[N-(carbamoylmethyl)pyrrolidin-3-yl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl-1-methyl-1-carbapen-2-em-3-carboxlic acid diastereomer A and B ##STR1214##

The same procedure as in Example 1-1) was carried out by using p-nitrobenzyl (1R,8S,6S)-2-diphenoxyphosphoryloxy-6-((R)-1-hydroxyethyl]-1-methyl-[-carbapen-2-em-3-carboxylate (570 mg, 0.96 mmol) and (2S,4S)-2-[N-(carbamoylmethyl)pyrrozidin-3-yl]-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine trifluoromethanesulfonate (540 mg, 0.97 mmol) to obtain p-nitrobenzyl (1R, 5S,6S)-2-[(2S,4S)-2[N-(carbamoylamethyl)pyrrolidin-3-yl]-N-(p-nitropenzyloxycarbonyl)pyrrolidin-2-ylthio]-6[(R]-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (197 mg, yield: 27.3%).

IR(KBr)cm⁻¹ : 1770, 1700, 1520, 1350

NMR(CDCl₃) δ: 1.28(3H,t,J=7Hz),1.35(3H,t,J=6Hz),3.61(1H,m),4.0-4.35(4H,m),5.10-5.34(3M,m),5.53(1H,d,J=14Hz),7.52 (1H,m),2.52-2.80(3H,m),2.97(2H,=,J=7Hz),3.03(1H,m), 3.22-3.64(7H,m),3.94(1H,m),4.24(2H,m)

HPLC(the same condition as in Example 1)

Retention time: 6.16 min

EXAMPLE 46 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2 -(pyrrolidin-2-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid diastereomer A ##STR1215##

The same procedure as in Example 39-1) was carried out by using (2S,4S)-4-acetylthio-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-2-yl)pyrrolidine diastereomer A (186 mg, 0,487 mmol, the compound of Reference Example 34) to obtain (2S,4S)-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-2-yl)-4-mercaptopyrrolidine diastereomer A.

The same procedure as in Example 1-1) was carried out by using thiol obtained by the above reaction and allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (243 mg, 0.487 mmol) to obtain allyl (1R,5S,6S)-2-[(2S, 4S)-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin- 2-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer A (160 mg, yield: 55.7% ).

IR (KBr)cm⁻¹ : 1780, 1700, 1550, 1410

NMR(CDCl₃) δ: 1.25(3H,d,J=7Hz),1.34(3H,d,J-6Hz),1.5-2.1(5H,m), 2.3-2.9(2H,m),3.07(1H,t,J=11Hz),3.2-3.8(6H,m),4.1-4.4(4H,m),4.5-5.0(6H,m),5.2-5.6(6H,m),5.8-6.2(3H,m) ##STR1216##

The same procedure as in Example 9-2) was carried out by using the compound obtained by the adore reaction (diastereomer A; 157 mg, 0.266 mmol) to obtain the above identified compound (diastereomer A; 28.5 mg, yield: 28%).

IR(KBr)cm⁻¹ : 1750, 1590, 1390

NMR(D₂ O) δ: 1.16(3H,d,J=7Hz),1.24(3B,d,J=6Hz),1.42(1H,m),1.74 (1H,m),2.00(2H,m),2.24(1H,m),2.53(1H,m),2.87(1H,dd, J=12,4Hz),3.1-3.6(7H,m),3.78(1H,m),4.1-4.3(2H,m)

HPLC(the same condition as in Example 1)

Retention time: 2.8 min

EXAMPLE 47 (1R,5S,6S)-6-[(R)-1-hydroxyl]-methyl-2-[(2S,4S)-2-(pyrrolidin-2-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid diastereomer B ##STR1217##

The same procedure as in Example 39-1) was carried out by using (2S,4S]-4-acetylthio-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-2-yl)pyrrolidine diastereomer B (133 mg, 0.348 mmol, the compound of Reference Example 35) to obtain (2S,4S)-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-2-yl)-4-mercaptopyrrolidine diastereomer B.

The same procedure as in Example 1-1) was carried out by using thiol obtained by the above reaction allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (174 mg, 0.348 mmol) to obtain allyl (1R,5S,6S)-2-[(2S, 4S)-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-2-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate diastereomer S (109 mg yield: 53.1%).

IR(KBr)cm⁻¹ : 1780, 1700, 1550, 1410

NMR(CDCl₃) δ: 1.26(3H,d,J=7Hz),1.35(3H,d,J=6Hz), 1.65(2H,m),1.93 (3H,m) ,2.3-2.7(2H,m),3.1-3.8(6H,m),4.0-4.5(4H,m), 4.5-3.0(6H,m),5.1-5.6(6H,m),5.8-6.1(3H,m) ##STR1218##

The same procedure as in Example 9-2) was carried out by using the compound obtained by the above reaction (diastereomer B; 103 mg, 0,175 mmol) to obtain the above identified compound (diastereomer B: 10.0 mg yield: 15%).

IR(KBr)cm⁻¹ : 1750, 1590, 1390

HPLC(the same condition as in Example 1)

Retention time: 3.4 min

EXAMPLE 48 (1R,5S,6S]-2-[(2S,4S)-2-(5-oxopiperazin-3-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid ##STR1219##

The same procedure as in Example 39-1) was carried out by using (2S,4S)-4-acetylthio-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-5-oxopiperazin-3-yl)pyrrolidine (168 mg, 0.44 mmol) to obtain (2S,4S)-N-allyloxycarbonyl-2-(1-allyloxycarponyl-5-oxopiperazin-3-yl)-4-mercaptopyrrolidine.

The same procedure as in Example 9-1) was carried out by using thiol obtained by the above reaction and allyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (200 mg, 0.40 mmol) to obtain allyl (1R,5S,6S)-2-[(2S, 4S)-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-5-oxo-piperazin-3-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (156 mg, yield: 58%).

NMR(CDCl.sub.) δ: 1.28(3H,d,J=8Hz),1.34(3H,d,J=8Hz),1.80(1H,m),2.60 (1H,m),2.90-5.00(19H,m),5.20-5.60(6H,m),5.96(3H,m) ##STR1220##

The same procedure as in Example 9-2) was carried out by using the compound obtained by the above reaction (156 mg, 0.25 mmol ) to obtain the above identified compound (50 mg, yield: 48% ).

NMR(D₂ O) δ: 1.24(3H,d,J 8Hz),1.30(3H,d,J=8Hz),1.76(1H,m),2.70 (1H,m),3.00(1H,m),3.10-4.10(10H,m),4.26(2H,m)

HPLC:

Eluent: 0.01M Phosphate buffer (pH 6.5)-Methanol (85:15)

Flow rate: 0.8 ml/min

Other conditions are the same as in Example 1

Retention time: 4.91 min

EXAMPLE 49 (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2[(2S,4S)-2-(N-methylpiperidin-4-yl)pyrrolidin-4-ylthio-1-carbapen-2-em-3-carboxylic acid ##STR1221##

To a solution of p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (2.50 g, 4.2 mmol) in acetonitrile (60ml), N,N-diisopropylethylamine was dropwise added in a nitrogen stream under cooling with ice, followed by dropwise adding a solution of (2S,4S)-4-mercapto-2-(N-methylpiperidin-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine trifluoroacetic acid salt (2.30 g, 4.6 mmol, compound of Reference Example 37 in acetonitrile (60 ml). After the mixture was stirred at 0° C. for 15 hours, the precipitate was collected by filtration, washed with acetonitrile and dried to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2 -[(2S,4S)-2-(N-methylpiperidin-4-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (2.12 g, yield: 69.7%).

IR(KBr)cm⁻¹ : 1770, 1700, 1520, 1345

NMR(CDCl₃) δ: 1.28(3H,d,J=8Hz),1.36(3H,d,J=7Hz),1.2-2.2(9H,m), 2.25(3H,s),2.2(1H,m),2.8-3.2(3H,m),3.2-3.6(3H,m), 3.97(1H,m),4.2-4.35(2H,m),5.15-5.3(3H,m),5.53(2H,d, J=13Hz),7.56(2H,d,J=9Hz),7.70(2H,d,J=8Hz),8.27(2H, d,J=8Hz),8.28(2H,d,J=9Hz) ##STR1222##

The same procedure as in Example 49-1) was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (3.86 g, 6.5 mmol), (2S,4S)-4-mercapto-2-(N-methylpiperidin-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine hydrochloride (2.88 g, 6.9 mmol, the compound of Reference Example 38 and N,N-diisopropylethylamine (2.61 ml, 15 mmol) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-1-[(2S,4S)-2-(N-methylpiperidin-4-yl)pyrrolidin-4-yl-thio]-1-carbapen-2-em-3-carboxylate (3.74 g, yield: 79.5%). Various spectra data of this compound agreed to the data of the compound obtained in Example 49-1). ##STR1223##

To a solution of the compound obtained by the above reaction (1.12 g, 1.55 mmol) in tetrahydrofuran 13 ml), ethanol (5 ml), a 0.25M sodium 3-morpholinopropanesulfonate buffer solution (pH7.0, 31 ml) and 10% palladium-carbon catalyst (750 mg) were added, and the mixture was stirred in hydrogen stream at room temperature for 3 hours. The catalyst was filtered off from the reaction mixture, and the organic solvent was distilled off under reduced pressure, and then insoluble matters in the aqueous layer were filtered off. The obtained filtrate was concentrated to 20 ml, and the residue was subjected to reversed phase column chromatography (YMC-GEL™ ODS-AQ 120-S50, a 20methanol aqueous solution). The desired fraction was concentrated and freeze-dried to obtain the above identified compound (556 mg, yield: 87.5%).

IR(KBr)cm⁻¹ : 1750, 1590, 1390

NMR(CDCl₃) δ: 1.21(3H,d,J=7Hz),1.29(3H,d,J=6Hz),1.2-1.82(6H,m), 1.84-2.16(2H,m),2.53(1H,m),2.74(3H,s),2.7-3.12(3H, m),3.12-3.5(4H,m),3.9(1H,m),4.13-4.32(2H,m)

EXAMPLE 50 (1R,5S,6S)-2-[(2S,4S)-2-(N-ethylpyrrolidin-3-yl]pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid ##STR1224##

Under a nitrogen atmosphere at -20 ° C., to a solution of p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (13.76 g, 23.1 mmol) in acetonitrile (150 ml) was added N,N-diisopropylethylamine (13.44 ml, 77.1 mmol), followed by a suspension of (2S,4S)-2-(N-ethylpyrrolidin-3-yl)-4-mercaptopyrrolidine dihydrochloride (7.0 g, 25.7 mmol, compound of Reference Example 40 in N,N-dimethylformamide (40 ml). After stirring overnight at -20° C., the reaction mixture was concentrated in vacuo. The resulting precipitate was collected by filtration, and washed with acetonitrile (30 ml) followed by isopropyl ether (30 ml) to obtain p-nitrobenzyl (1R,5S,6S)-2-[(2S, 4S)-2-(N-ethylpyrrolidin-3-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (10.1 g, yield: 72%) as a crude crystal.

IR(KBr)cm⁻¹ : 3350, 2900, 2660, 1770, 1700, 1605, 1515

NMR(DMSO-d₆) δ: 1.18(3H,d,J=8Hz),1.19(3H,d,J=8Hz),1.24(3H,t,J=9Hz), 1.6(2H,m),1.8-2.3(2H,m),2.4-3.8(12H,m),4.0(1H,m), 3.0(1H,m),5.15(1H,m),5.28(1H,d,J=16Hz),5.5(1H,d,J=16Hz),7.75(2H,d,J=8Hz),8.27(2H,d,J=8Hz)

mp. 216 ° C. (dec) ##STR1225##

To a solution of the compound obtained by the above reaction (10 g, 18.3 mmol) in tetrahydrofuran (500 ml), 0.5 M 3-morpholinopropanesulfonic acid buffer (pH 7.0, 500 ml) and ethanol (150 ml) were added 10% palladium-carbon catalyst (10 g), and the mixture was hydrogenated at atmospheric pressure and room temperature for 2 hours. The catalyst was removed by filtration, and the filtrate was concentrated in order to remove tetrahydrofuran and ethanol in vacuo. The residual aqueous solution was washed with chloroform, and a small amount of insoluble matter was removed by filtration. The resulting aqueous solution was again concentrated in vacuo, and purified by column chromatography (HP-20SS, 250 ml, elution with 30% aqueous methanol) to obtain the above identified compound (3.39 g, yield: 45.2%) as a crystal. The mother Liquor was again purified to obtain the second crystal (1.94 g, yield: 25.9%).

IR(KBr)cm⁻¹ : 3220, 1745, 1675, 1580, 1540, 1390

NMR(DMSO-d₆) δ: 1.18(3H,d,J=8Hz),1.19(3H,d,J=8Hz),1.24(3H,t,J=9Hz), 1.2-1.8(2H,m),2.2-3.0(6H,m),3.0-3.2(2H,m),3.4-4.2 (10H,m)

UV λ_(max) (0.1 M 3-morpholinopropanesulfonic acid buffer, pH 7.0):

299 nm (ε=6840)

EXAMPLE 51 (1R,5S,6S)-2-[(2S,4S)T2-(N-ethylpiperidin-4-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid ##STR1226##

The same procedure as in Example 49-1) was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (595 mg, 1.0 mmol), (2S,4S)-2-(N-ethylpiperidin- 4-yl)-4-mercapto-N-p-nitrobenzyloxycarbonyl)pyrrolidine hydrochloride (463 mg, 1.08 mmol, synthesized by the same procedure as in Reference Example 37), N,N-diisopropylethylamine (0.4 ml, 2.3 mmol) and acetonitrile (28 ml) to obtain p-nitrobenzyl (1R,5S,6S)-2-[(2S,4S)-2-(N-ethylpiperidin-4-yl)-(N-p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (479 mg, yield: 64.9%).

IR(KBr)cm⁻¹ : 1770, 1700, 1520, 1345 ##STR1227##

The same procedure as in Example 49-3) was carried out by using the compound obtained by the above reaction (460 mg, 0.62 mmol), tetrahydrofuran (12 ml), ethanol (2 ml), 0.25 M sodium 3-morpholinopropanesulfonate buffer solution (pH 7.0, 12 ml) and 10% palladium-carbon catalyst (308 mg) to obtain the above identified compound (189 mg, yield: 72.3%).

IR(KBr)cm⁻¹ : 1750, 1590, 1390

EXAMPLE 52 (1R,5S,6S)-2-[(2S,4S),2-(N,N-dimethyl-4-piperidinio)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate ##STR1228##

To a solution of p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methylpiperidin-4-yl)pyrrolidin-4-ylthio]-! -carbapen-2-em-3-carboxylate (1.0 g, 1.38 mmol, the compound of Example 49-1) in acetone (25 ml), methyl iodide (0.86 ml, 13.8 mmol) was added. The mixture was stirred at room temperature for 15 hours, and the organic solvent was distilled off under reduced pressure.

The same procedure as in Example 49-3) was carried out by using the obtained residue, tetrahydrofuran (20 ml), ethanol (20 ml), 0.25 M sodium 3-morpholinopropanesulfonate buffer solution (pH 7.0, 31 ml) and 10% palladium-carbon catalyst (750 mg) to obtain the above identified compound (402 mg, yield: 69.3%).

IR(KBr)cm⁻¹ : 1750, 1590, 1390

NMR(D₂ O) δ: 1.21(3H,d,J=7Hz),1.29(3H,d,J=6Hz),1.2-1.5(2H,m), 1.63-2.15(6H,m),2.54(1H,m),3.08(3H,s),3.15(3H,s), 2.84-3.6(7H,m),3.78(1H,m),4.13-4.33(2H,m)

EXAMPLE 53 (1R,5S6S)-2-[(2S,4S),2-(azetidin-3-yl)pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid ##STR1229##

The same procedure as in Example 1-1) was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (83 mg, 0.14 mmol) and (2S,4S)-4- mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)azetidin-3-yl]pyrrolidine (73 mg, 0.14 mmol, compound of Reference Example 41) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-[N-(p-nitrobenzyloxycarbonyl)azetidin-3-yl]pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (86.8 mg, yield: 72.2%).

IR(KBr)cm⁻¹ : 1770, 1700, 1600, 1520, 1340

NMR(CDCl₃) δ: 1.28(3H,d,J=8Hz),1.35(3H,d,J=7Hz),1.72(1H,m),2.32-2.72(2H,m),2.95-3.50(4H,m),5.10-5.34(5H,m),5.53(1H, d,J=15Hz),7.51(4H,d,J=9Hz),1.66(2H,d,J=9Hz),8.22 (6H,d,J=9Hz) ##STR1230##

The same procedure as in Example 1-2) was carried out by using the compound obtained by the above reaction (86.8 mg, 0.10 mmol) to obtain the above identified compound (14.4 mg, yield: 38.8%).

IR(KBr)cm⁻¹ : 1750, 1600, 1390

NMR(D₂ O) δ: 1.18(3H,d,J=8Hz),1.27(3H,d,J=7Hz),2.48(1H,m),2.88 (1H,dd,J=14,4Hz),3.03-3.30(3H,m),3.32-3.57(3H,m), 3.76(1H,m),3.95(2H,m),4.20(4H,m)

EXAMPLE 54 (1R,5S,6S)-6-[(R)-1-hydroxyethyl-1-methyl-2-[(2S,4S)-2-(N-methylazetidin-3-yl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylic acid ##STR1231##

The same procedure as in Example 49-1) was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (300 mg, 0.50 mmol) and (2S,4S)-4-mercapto-2-(N-methylazetidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine hydrochloride (195 mg, 0.50 mmol, compound of Reference Example 42) to obtain p-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methylazetidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (280 mg, yield: 80% ).

IR(KBr)cm⁻¹ : 1770, 1700, 1520, 1350 ##STR1232##

The same procedure as in Example 49-2) was carried out by using the compound obtained by the above reaction (140 mg, 0.20 mmol) to obtain the above identified compound 42 mg, yield: 55%). IR(KBr)cm⁻¹ : 1750, 1590, 1390

EXAMPLE 55 (1R,5S,6S)-2-[(2S,4S)-2-(N,N-dimethyl-3-azetidinio)pyrrolidin-4-ythio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate ##STR1233##

The same procedure as in Example 52 was carried out by using o-nitrobenzyl (1R,5S,6S)-6-[(R)-1-hydroxyethyl]-1-methyl-2-[(2S,4S)-2-(N-methylazetidin-3-yl)-N-p-nitropenzyloxycarbonyl)pyrrolidin-4-ylthio]-1-carbapen-2-em-3-carboxylate (140 mg, 0.20 mmol, compound of Example 54-1) to obtain the above identified compound (32 mg, yield: 40.6%).

IR(KBr)cm⁻¹ : 1750, 1590, 1390

EXAMPLE 56 (1R,5S,6S)-2-[(2S,4S)-2-(N-ethylazetidin-3-yl(pyrrolidin-4-ylthio]-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylic acid ##STR1234##

The same procedure as in Example 49-1) was carried out by using p-nitrobenzyl (1R,5S,6S)-2-diphenoxyphosphoryloxy-6-[(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (168 mg, 0.28 mmol) and (2S,4S)-2-(N-ethylazetidin-3-yl)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine hydrochloride (113 mg, 0.28 mmol, compound of Reference Example 43) to obtain p-nitrobenzyl (1R,5S,6S)-2-[(2S,4S)-2-(N-ethylazetidin-3-yl)-1)-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio]-6-(R)-1-hydroxyethyl]-1-methyl-1-carbapen-2-em-3-carboxylate (130 mg, yield: 65%).

IR(KBr)cm⁻¹ : 1770, 1700, 1520, 1350 ##STR1235##

The same procedure as in Example 49-3) was carried out by using the compound obtained by the above reaction (130 mg, 0.18 mmol) to obtain the above identified compound (38 mg, yield: 52.4%).

IR(KBr)cm⁻¹ : 1750, 1590, 1390

REFERENCE EXAMPLE 1 (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidin-4-yl)pyrrolidine ##STR1236##

To an aqueous solution (40 ml) of L-hydroxyproline methyl ester hydrochloride (8.2 g, 45.2 mmol), triethylamine (7.54 ml, 54.2 mmol) was added at room temperature, and then a solution of 2-(tert-butoxycarbonylthio)-4,6-dimethylpyrimidine (10.8 g, 45 mmol) in dioxane (80 ml) was added thereto. The reaction solution was stirred at room temperature for 1.5 hours, and then dioxane was distilled off under reduced pressure. The residue was extracted with ethyl acetate (250 ml). The organic layer was washed sequentially with a 0.1 N sodium hydroxide aqueous solution, dilute hydrochloric acid, water and a saturated sodium chloride aqueous solution and then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 5% methanol-methylene chloride) to obtain N-tert-butoxycarbonyl-L-hydroxyproline methyl ester (9.3 g, yield: 84%).

NMR(CDCl₃) δ: 1.38-1.48(9H,m) ,1.98-2.38(2H,m) ,3.40-3.72 (2H,m), 3.86 (3H,s),4.35-4.58(2H,m) ##STR1237##

Imidazole (4.95 g, 72.7 mmol) and tert-butyldimethylsilyl chloride (10.7 g, 71.0 mmol) were added to a solution of the compound obtained by the above reaction (14.0 g, 57.1 mmol) in N,N-dimethylformamide (150 ml) in a nitrogen stream at room temperature. This solution was stirred overnight at room temperature. Ethyl acetate (500 ml) was added to the reaction solution, and the organic layer was washed twice with water and once with a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-L-proline methyl ester (19 g, yield: 92.6%). ##STR1238##

To a solution of the compound obtained by the above reaction (19.0 g, 52.9 mmol) in tetrahydrofuran (180 ml), lithium chloride (4.49 g, 106 mmol) and then sodium borohydride (4.0 g, 106 mmol) were added in a nitrogen stream. To this mixture, ethanol (180 ml) was dropwise added, and the reaction solution was stirred overnight at room temperature. A saturated ammonium chloride aqueous solution (100 ml) was added to this reaction mixture to decompose an excess reducing agent. Then, this mixture was concentrated under reduced pressure. The residue was extracted with ethyl acetate (500 ml), and the organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 5:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-hydroxymethylpyrrolidine (14.88 g, yield: 84.9%). ##STR1239##

A solution of dimethyl sulfoxide (3.6 ml, 50.7 mmol) in methylene chloride (15 ml) was dropwise added to a solution of oxalyl chloride (2.89 ml, 33.9 mmol) in methylene chloride (100 ml) in a nitrogen stream at -78° C. The reaction solution was stirred at the same temperature for 30 minutes. To this mixture, a solution of (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-hydroxymethylpyrrolidine (8.0 g, 24.2 mmol) in methylene chloride (50 ml) was dropwise added at -78° C. This mixture was stirred at the same temperature for 30 minutes, and then triethylamine (11.1 ml, 79.8 mmol) was added thereto. The mixture was stirred further for 30 minutes. The reaction mixture was poured into methylene chloride (200 ml), and the organic layer was washed sequentially with dilute hyrochloric acid, water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain an oily residue of (2S,4 R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-formylpyrrolidine.

Ethyl diethoxyphosphorylacetate (6.0 g, 26. 8 was added to a suspension of 60% sodium hydride (2.0 g, 25.0 mmol) in tetrahydrofuran (100 ml) in a nitrogen stream under cooling with ice. This solution was stirred at the same temperature for 30 minutes. To this reaction solution, a tetrahydrofuran solution (20 ml) of (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-formylpyrrolidine obtained by the above reaction, was dropwise added, and the mixture was stirred further at the same temperature for 30 minutes. The reaction solution was extracted with ethyl acetate (150 ml). The organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 10:1) to obtain (E)-3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2yl])-acrylic acid ethyl ester (9.4 g, yield: 97.5%).

NMR(CDCl₃) δ: 0.06(6H,s),0.88(9H,s),1.30(3H,d,J=7Hz),1.44 (9H,br s),1.84(1H,m),2.10(1H,m),3.48(2H,m),4.12(2H,q, J=7Hz),4.35(1H,m),5.88(1H,br d,J=16Hz),6.86(1H,m) ##STR1240##

1,1,3,3-tetramethylguanidine (5.8 ml, 46.2 mmol) was dropwise added to a solution of the compound obtained by the above reaction (9.0 g, 22.6 mmol) in nitromethane (33 ml) in a nitrogen stream at room temperature. The mixture was stirred overnight at the same temperature. Ethyl acetate (200 ml) was added to the reaction solution, and the organic layer was washed sequentially with dilute hydrochloric acid, water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 10:1) to obtain 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-buthyldimethylsiloxypyrrolidin-2-yl]-4nitrobutyric acid ethyl ester (10.1 g, yield: 97.3%).

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.27(3H,d,J=7Hz),1.48 (9H,br s),1.73(1H,m),2.00(1H,m),2.44(2H,m),3.16 (1H,m),4.16(4H,m),4.33(1H,m)4.53(1H,m) ##STR1241##

Raney nickel (W-2 type, 3.0 ml) was added to a solution of the compound obtained by the above reaction (3.6 g, 7.8 mmol) in ethanol (50 ml). The mixture was stirred overnight at room temperature in a hydrogen atmosphere. The catalyst was filtered off from the reaction solution. The filtrate was concentrated under reduced pressure. The residue was dissolved in benzene (50 ml) and the mixture was refluxed overnight. The reaction solution was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (Wakogel™ C-300, 1% methanol-chloroform) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-pyrrolidon-4-yl)pyrrolidine (1.87 g, yield: 62.2%).

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.47(9H,s),1.72(1H,m),1.88-2.48 (3H,m),2.95-3.75(5H,m),4.13(1H,m),4.30(1H,m) ##STR1242##

The compound obtained by the above reaction (1.87 g, 4.87 mmol) was dissolved in a 80% trifluoroacetic acid aqueous solution (20 ml), and the solution was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in a mixture comprising dioxane (10 ml) and water (10 ml). The pH was adjusted to 8 with a 1N sodium hydroxide aqueous solution. Then, 4,6-dimethyl-2-(p-nitrobenzyloxycarbonylthio)pyrimidine (1.55 g, 4.86 mmol) was added thereto. This solution was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was extracted with ethyl acetate (100 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 3% methanol-chloroform) to obtain (2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine (769 mg, yield: 45.2%).

NMR(CDCl₃) δ: 1.72-2.52(5H,m) ,2.98-3.58(3H,m) ,3.78(1H,m), 4.24(1H,m),4.48(1H,m),5.25(2H,br s),7.54 (2H,d,J=9Hz),8.24(2H,d,J=9Hz) ##STR1243##

Triphenylphosphine (567 mg, 2.16 mmol) was added to a solution of the compound obtained by the above reaction (302 mg, 0.87 mmol) in tetrahydrofuran (10 ml) under a nitrogen stream, and then diethyl azodicarboxylate (0.341 ml, 2.17 mmol) was dropwise added thereto under cooling with ice. The reaction solution was stirred for 30 minutes under cooling with ice. Then, thioacetic acid (0.155 ml, 2.17 mmol) was dropwise added thereto. The mixture was stirred at room temperature for two hours. Then, the reaction solution was extracted with ethyl acetate (100 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300) to obtain (2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)- 2-(2-pyrrolidon-4-yl)pyrrolidine (166 mg, yield: 47.1%) by the elution with hexane-ethyl acetate (1:1) and (2R,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine (166 mg, yield: 47.1%) by the elution with 3% methanol-chloroform.

(2S,4S) isomer

1R(KBr)cm⁻¹ : 1700, 1520, 1340, 1110

NMR(CDCl₃) δ: 1.70(1H,m),1.98-2.62(4H,m),2.37(3H,s),3.17(2H,m),3.42 (1H,m),3.87(1H,m),4.05-4.35(2H,m),5.24 (2H,br s),7.55(2H,d,J=9Hz),8.26(2H,d,J=9Hz)

(2R,4S) isomer

1R(KBr)cm⁻¹ : 1700, 1520, 1350, 1120

NMR(CDCl₃) δ: 1.68(1H,m),2.30-2.95(4H,m),2.38(3H,s),3.20(1H,m),3.55 (1H,m),3.88(2H,m),4.06-4.30(2H,m),5.26(2H,br s), 7.56(2H,d,J=9Hz),8.26(2H,d,J=9Hz) ##STR1244##

A 1N sodium hydroxide aqueous solution (0.42 ml) was dropwise added to a solution of the (2S,4S) isomer obtained by the above reaction (166 mg, 0.41 mmol) in methanol (10 ml) in a nitrogen stream under cooling with ice. This solution was stirred at the same temperature for 15 minutes. To the reaction solution, 1N hydrochloric acid (0.45 ml) was dropwise added. Then, the reaction solution was concentrated under reduced pressure. The residue was extracted with ethyl acetate (70 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine (140 mg, yield: 94%).

REFERENCE EXAMPLE 2 (2R,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine ##STR1245##

The same procedure as in Reference Example 1-9 was carried out by using (2R,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine (166 mg, 0.41 mmol, compound of Reference Example 1-8) to obtain the above identified compound (140 mg, yield: 94%).

REFERENCE EXAMPLE 3 (2S,4S)-2-(2-azetidinon-4-yl)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer ##STR1246##

Benzylamine (1.2 ml, 11 mmol) was added to [(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]acrylic acid ethyl ester (2.18 g, 5.46 mmol, compound of Reference Example 1-4). The mixture was stirred at room temperature for five days. Then, this mixture was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 3:1) to obtain 3-benzylamino-3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]propionic acid ethyl ester (1.95 g, yield: 70.5%). NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,s),1.24(3H,t,J=8Hz),1.44(9H,s), 1.90(1H,m),2.35(2H,m),3.26(1H,m),3.89 and 3.96 (2H,ABq,J=8Hz),4.14(2H,q,J=8Hz),4.39(1H,m), 7.31(5H,m) ##STR1247##

A 1N sodium hydroxide aqueous solution (4.3 ml) was dropwise added to a solution of the compound obtained by the above reaction (1.98 g, 3.9 mmol) in ethanol (30 ml) at room temperature. This solution was stirred overnight at the same temperature. 1N hydrochloric acid (4.3 ml) was added to this reaction solution. This solution was concentrated under reduced pressure. Tetrahydrofuran (50 ml) was added to the residue, and insoluble matters were filtered off. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The obtained oily substance (1.83 g) was dissolved in acetonitrile (380 ml). Triphenylphosphine (1.2 g, 4.58 mmol) and 2,2'-dipyridyl disulfide (1.01 g, 4.58 mmol) were added thereto in a nitrogen stream. This solution was stirred at 80° C. for 4.5 hours. The reaction solution was concentrated under reduced pressure and extracted with ethyl acetate (150 ml). The organic layer was washed sequentially with a 0.1N sodium hydroxide aqueous solution, water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 10:1→3:1) to obtain (2S,4R)-2-(N-benzyl-2-azetidinon-4-yl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidine diastereomer B (1.07 g, yield: 60.7%, highly polar compound) and diastereomer A (0.47 g, yield: 26.8%).

Diastereomer B

1R(KBr)cm⁻¹ : 1730, 1690, 1390, 1170

NMR(CDCl₃) δ: 0.06(6H,s),0.88(9H,s),1.50(9H,br s),2.54(1H,br d, J=14Hz),2.93(1H,dd,J=14,4Hz),3.16(1H,dd,J=12,4Hz), 7.35(5H,m)

Diastereomer A

1R(KBr)cm⁻¹ : 1760, 1700, 1400, 1260

NMR(CDCl₃) δ: 0.04(6H,s),0.85(9H,s),1.45(9H,br s),1.84(2H,m), 2.58(1H,br d,J=16Hz),2.92(2H,m),3.44(1H,m),3.95-4.30 (4H,m),4.60(1H,m),7.30(5H,m) ##STR1248##

Liquid ammonia (about 30 ml) was added to a solution of the diastereomer B obtained in the above reaction (470 mg, 0.98 mmol) in tetrahydrofuran (10 ml) and tert-butyl alcohol (1 ml) at -78° C. Sodium metal (100 mg, 4.35 mmol) was added to the reaction solution at the same temperature. The mixture was stirred for 15 minutes, and ammonium chloride (465 mg, 8.69 mmol) was added thereto. The reaction mixture was returned to room temperature, and ammonia was distilled off. The residue was extracted with ethyl acetate (100 ml). The organic layer was washed sequentially with water and a saturated sodium chloride solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 1:1) to obtain (2S,4R)-2-(2-azetidinon-4-yl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidine diastereomer B (340 mg, yield: 89.1%).

1R(KBr)cm⁻¹ : 1750, 1700, 1380, 1260

NMR(CDCl₃) δ: 0.06(6H,s),0.87(9H,s),1.47(9H,s),1.61(1H,m),2.02 (1H,m),2.62(1H,br d,J=16Hz),3.02(1H,dd,J=16,6Hz), 3.33(1H,m),3.50(2H,m),4.05(1H,m),4.30(1H,m) ##STR1249##

Anisole (one drop) and trifluoroacetic acid (3 ml) were dropwise added to a solution of the compound obtained by the above reaction (340 mg, 0.88 mmol) in methylene chloride (3 ml) in a nitrogen stream under cooling with ice. The reaction solution was stirred for one hour under cooling with ice and concentrated under reduced pressure. The residue was dissolved in tetrahydrofuran (5 ml). Triethylamine (1.2 ml, 8.6 mmol) and 4,5-dimethyl-2-(p-nitrobenzyloxycarbonylthic)pyrimidine (280 mg, 0.88 mmol) were added thereto under cooling with ice, and the mixture was stirred at room temperature for 1.5 hours. The reaction solution was extracted with ethyl acetate (70 ml). The organic layer was washed sequentially with a 0.1N sodium hydroxide aqueous solution, water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was dissolved in acetonitrile (5 ml), and 45% hydrofluoric acid (0.5 ml) was dropwise added thereto at room temperature. The mixture was stirred at room temperature for 1.5 hours. Then, the reaction solution was extracted with ethyl acetate (70 ml). The organic layer was washed sequentially with a 5% sodium hydrogen carbonate solution, water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 3% methanol-chloroform) to obtain (2S,4R)-2-(2-azetidinon-4-yl)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer B (177 mg, yield: 60.3%).

IR(KBr)cm⁻¹ : 1750, 1700, 1520, 1340

NMR(CDCl₃) δ: 1.72(1H,m),2.14(1H,m),2.64(1H,d,J=14Hz),3.06(2H,m), 3.57(1H,m),3.74(1H,m),4.12(1H,m),4.44(1H,m), 5.22(2H,br s),7.52(2H,d,J=8Hz),8.22(2H,d,J=8Hz) ##STR1250##

Triphenylphosphine (347mg, 1.32 mmol) was added to a solution of the compound obtained by the above reaction (177 mg, 0.53 mmol) in tetrahydrofuran (10 ml) in a nitrogen stream under cooling with ice, and then diethyl azodicarboxylate (0.208 ml, 0,132 mmol) was dropwise added thereto. The reaction solution was stirred for 30 minutes under cooling with ice, and then thioacetic acid (95 μl, 0,132 mmol) was dropwise added thereto. This reaction mixture was stirred at the same temperature for 3 hours. Then, the reaction solution was extracted with ethyl acetate (50 ml). The organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate) to obtain (2S,4S)-4-acetylthio-2-(2-azetidinon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer B (77 mg, yield: 37.1%).

NMR(CDCl₃) δ: 1.65(1H,m),2.38(3H,s),3.08(1H,dd,J=16,4Hz),3.30(1H,t, J=9Hz),3.64-4.28(4H,m),5.25(2H,br s),7.55 (2H,d,J=8Hz),8.26(2H,d,J=8Hz) ##STR1251##

A 0.1N sodium hydroxide aqueous Solution (0.196 ml) was dropwise added to a solution of the compound obtained by the above reaction (77 mg, 0.22 mmol) in methanol (10 ml) in a nitrogen stream under cooling with ice. The reaction solution was stirred for 15 minutes under cooling with ice, and then 1N hydrochloric acid (0.21 ml) was added thereto. This reaction mixture was concentrated under reduced pressure. The residue was extracted with ethyl acetate (50 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain (2S,4S)-2-(2-azetidinon-4-yl)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer B (67 mg, yield: 97%).

REFERENCE EXAMPLE 4 (2S,4S)-2-(2-azetidinon-4-yl)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer A ##STR1252##

The same procedure as in Reference Example 3-3 was carried out by using (2S,4R)-2-(N-benzyl-2-azetidinon-4-yl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidine diastereomer A (500 mg, 1.05 mmol) and sodium metal (75 mg, 3.26 mmol) to obtain (2S,4R)-2-(2-azetidinon-4-yl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidine diastereomer A (360 mg, yield: 88.4%).

1R(KBr)cm⁻¹ : 1760, 1740, 1690, 1390, 1260

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.47(9H,s),1.93(2H,m),2.66 (1H,d,J=16Hz),2.97(1H,ddd,J=16,5,2Hz),3.26 (1H,dd,J=(2,5Hz),3.55(1H,m),4.00-4.44(3H,m) ##STR1253##

The same procedure as in Reference Example 3-4 was carried out by using the compound obtained by the above reaction (350 mg, 0.90 mmol) to obtain (2S,4R)-2-(2-azetidinon-4-yl)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer A (185 mg, yield: 61%).

1R(KBr)cm⁻¹ : 1740, 1670, 1520, 1440, 1340

NMR(CDCl₃) δ: 2.06(2H,m),2.70(1H,d,J=16Hz),3.02(1H,br d,J=16Hz), 3.47(1H,m),3.76(1H,m),4.16(1H,m),4.34(1H,m),4.50 (1H,m),5.26(2H,br s),7.54(2H,d,J=8Hz),8.24 (2H,d,J =8Hz) ##STR1254##

The same procedure as in Reference Example 3-5 was carried out by using the compound obtained by the above reaction (185 mg, 0.55 mmol), diethyl azodicarboxylate (0.218 ml, 0.138 mmol), triphenylphosphine (365 mg, 0.138 mmol) and thioacetic acid (99/μl, 0.138 mmol) to obtain (2S,4S)-4-acetylthio-2-(2-azetidinon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer A (107 mg, yield: 49.3% ).

NMR(CDCl₃) δ: 1.87 (1H,m),2.36(3H,s),2.52(1H,m),2.78(1H,m), 3.04(1H,br d,J=12Hz),3.18(1H,t,J=10Hz),3.80-4.32(4H,m),5.24(2H,br s),7.54(2H,d,J=8Hz), 8.23(2H,d,J=8Hz) ##STR1255##

The same procedure as in Reference Example 3-6 was carried out by using the compound obtained by the above reaction (107 mg, 0.27 mmol) and a 1N sodium hydroxide aqueous solution (0.27 ml) to obtain (2S,4S)-2-(2-azetidinon-4-yl)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer A (90.8 mg, yield: 95%).

REFERENCE EXAMPLE 5 (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidine ##STR1256##

Triethylamine (0.53 ml, 3.81 mmol), 4-dimethylaminopyridine (465 mg, 3.81 mmol) and di-tert-butyl dicarbonate (1.66 g, 760 mmol) were added to a solution of the compound obtained in Reference Example 1-6 (1.46 g, 3.80 mmol) in methylene chloride (15 ml) in a nitrogen stream at room temperature. The mixture was stirred at the same temperature for 4.5 hours. The reaction solution was concentrated under reduced pressure, and the residue was extracted with ethyl acetate (50 ml). The organic layer was washed with dilute hydrochloric acid, water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 5:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonyl-2-pyrrolidon-4-yl)pyrrolidine (1.76 g, yield 96%).

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.48(9H,s),1.54(9H,s),1.68 (1H,m),2.00(1H,m),3.25(1H,m),4.10(1H,m),4.34(1H,m) ##STR1257##

A borane-dimethyl sulfide complex (1.09 ml, 10.9 mmol) was dropwise added to a solution of the comound obtained by the above reaction (1.76 g, 3.64 mmol) in tetrahydrofuran (18 ml) in a nitrogen stream at room temperature, and the mixture was then refluxed for 1.5 hours. Methanol (5 ml) was added to the reaction solution under cooling with ice and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 10:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonylpyrrolidin-3-yl)pyrrolidine (1.62 g, yield: 94.8%).

NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,s),1.46(18H,s),1.60-2.04(4H,m), 4.07(1H,m),4.34(1H,m) ##STR1258##

A solution of 1.6N hydrogen chloride in methanol (15 ml) was added to the compound obtained by the above reaction (1.60 g, 3.40 mmol), and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure. The residue was dissolved in a mixture of dioxane (20 ml) and water (10 ml), and the solution was adjusted to pH 8.5 with triethylamine. Then, 4,6-dimethyl-2-(p-nitrobenzyloxycarbonylthio)pyrimidine (2.17 g, 6.80 mmol) was added thereto. This reaction solution was stirred at room temperature for 1.5 hours. The reaction solution was concentrated under reduced pressure, and the residue was extracted with ethyl acetate (50 ml). The organic layer was washed with a 1N sodium hydroxide aqueous solution, water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 2% methanol-chloroform) to obtain (2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidine (1.55 g, yield: 88.6%).

NMR(CDCl₃) δ: 1.52-2.20(4H,m),3.76(1H,m),4.23(1H,m),4.48(1H,m),5.22 (4H,s),7.52(4H,d,J=8Hz),8.20(4H,d,J=8Hz) ##STR1259##

Triethylamine (0.52 ml, 3.74 mmol) and methanesulfonyl chloride (0.28 ml, 3.62 mmol) were added to a solution of the compound obtained by the above reaction (1.55 g, 3.02 mmol) in tetrahydrofuran (25 ml) in a nitrogen stream under cooling with ice, and the mixture was stirred at the same temperature for 30 minutes. The reaction solution was extracted with ethyl acetate (50 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate) to obtain (2S,4R)-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidine (1.76 g, yield 98.6%).

NMR(CDCl₃) δ: 3.04(3H,s),4.18(2H,m),5.24(4H,br s),7.53(4H,d,J=8Hz), 8.24 (4H, d, J=8Hz ) ##STR1260##

Thioacetic acid (0.31 ml, 4.50 mmol) was dropwise added to a solution of anhydrous potassium carbonate (616 mg, 4.46 mmol) in N,N-dimethylformamide (30 ml) in a nitrogen stream at 0° C., and the mixture was stirred at the same temperature for 20 minutes. To this mixture, a solution of the compound obtained by the above reaction (1.76 g, 2.97mmol) in N,N-dimethylformamide (5 ml) and sodium iodide (450 mg, 30 mmol) were added at 0° C. This reaction mixture was stirred overnight at a temperature of from 60° to 70° . The reaction mixture was extracted with ethyl acetate (100 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 1:1) to obtain (2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-3-yl]pyrrolidine (1.34 g, yield: 78.8%).

NMR(CDCl₃) δ: 1.54-2.10(4H,m),2.36(3H,s),3.90(1H,m),4.12(1H,m),4.26 (1H,m),5.24(4H,s),7.52(4H,d,J=8Hz),8.24(4H,d,J=8Hz) ##STR1261##

The compound obtained by the above reaction (1.34 g, 2.34 mmol) was dissolved in a mixture of methanol (25 ml) and tetrahydrofuran (10 ml). A 1N sodium hydroxide aqueous solution (2.46 ml) was dropwise added thereto in a nitrogen stream under cooling with ice, and the mixture was stirred at the same temperature for 15 minutes. IN hydrochloric acid (2.53 ml) was dropwise added to the reaction solution, and the mixture was concentrated under reduced pressure. The residue was extracted with ethyl acetate (50 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain the above identified compound (1.24 g, yield: 100%).

REFERENCE EXAMPLE 6 (2S,4S)-4-mercapto-2-(N-methylpyrrolidin-3-yl)-p-nitrobenzyloxycarbonyl)pyrrolidine trifluoromethane-sulfonate ##STR1262##

2,6-lutidine (1.04 ml, 8.93 mmol) and trimethylsilyl trifluoromethanesulfonate (1.34 ml, 6.70 mmol) were added to a solution of the compound obtained in Reference Example 1-6 (1.72 g, 4.48 mmol) in methylene chloride (17 ml) in a nitrogen stream at room temperature, and the mixture was stirred at the same temperature for 30 minutes. Methanol (5 ml) was dropwise added to the reaction solution, and then the solution was concentrated. Dioxane (15 ml) was added to the residue. Triethylamine (0.94 ml, 6.72 mmol) and 4,6-dimethyl-2-(p-nitrobenzyloxycarbonylthio)pyrimidine (1.43 g, 4.48 mmol) were added to this reaction solution, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was extracted with ethyl acetate (50 ml). The organic layer was washed with dilute hydrochloric acid, water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate) to obtain (2S,4R)-4-tert-butyldimethylsiloxy-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine (813 mg, yield: 39.2%).

NMR(CDCl₃) δ: 0.03 (3H,s),0.06(3H,s),0.84(9H,s),1.78(1H,m), 1.92-2.50(3H,m),2.93-3.54(4H,m),3.66(1H,m),4.10 (1H,m),4.38(1H,m),5.23(2H,ABq,J=14Hz),7.52 (2H,d,J=8Hz),8.23(2H,d,J=8Hz) ##STR1263##

The same procedure as in Reference Example 5-1 was carried out by using the comopund obtained by the above reaction (813 mg, 1.76 mmol) and di-tert-butyl dicarbonate (766 mg, 3.51 mmol) to obtain (2S,4R)-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonyl-2-pyrrolidon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (944 mg, yield: 95.5%).

NMR(CDCl₃) δ: 0.04(3H,s),0.06(3H,s),0.84(9H,s),1.52(9H,s), 1.74(1H,m),2.00(1H,m),2.20-2.96(3H,m),4.19(1H,m), 4.39(1H,br s),5.24(2H,ABq,J=14Hz),7.53(2H,d,J=8Hz), 8.24(2H,d,J=8Hz) ##STR1264##

The same procedure as in Reference Example 5-2 was carried out by using the compound obtained by the above reaction (944 mg, 1.68 mmol) and a borane-dimethyl sulfide complex (0.50 ml, 5.0 mmol) to obtain (2S,4R)-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonylpyrrolidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (730 mg, yield: 79.3%).

NMR(CDCl₃₃) δ: 0.05(3H,s),0.07(3H,s),0.85(9H,s),1.45(9H,s), 1.52-2.08(4H,m),2.82(1H,m),2.90-3.80(6H,m), 4.15(1H,m),4.40(1H,br s),5.24(2H,m), 7.54(2H,d,J=8Hz),8.23(2H,d,J=8Hz) ##STR1265##

A solution of 1M tetrabutylammonium fluoride in tetrahydrofuran (13.7 ml) was added to a solution of the compound obtained by the above reaction (6.98 g, 12.7 mmol) in tetrahydrofuran (100 ml) in a nitrogen stream under cooling with ice, and the mixture was stirred at the same temperature for 40 minutes. A saturated ammonium chloride aqueous solution (20 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (150 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was dissolved in tetrahydrofuran (100 ml). Then, triethylamine (1.77 ml, 12.7 mmol) and methanesulfonyl chloride (0.99 ml, 12.8 mmol) were added thereto under cooling with ice, and the mixture was stirred at the same temperature for 1 hour. The reaction solution was extracted with ethyl acetate (150 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 1:1) to obtain (2S,4R)-2-(N-tert-butoxycarbonylpyrrolidin-3-yl)-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (6.64 g, yield: 100%).

NMR(CDCl₃) δ: 1.36(9H,s),2.96(3H,s),3.98-4.24(2H,m),5.18(2H,m),7.46 (2H,d,J=8Hz),8.16(2H,d,J=8Hz) ##STR1266##

The same procedure as in Reference Example 5-5 was carried out by using the compound obtained by the above reaction (6.6 g, 12.9 mmol) and thioacetic acid (1.33 ml, 19.3 mmol) to obtain (2S,4S)-4-acetylthio-2-(N-tert-butoxycarbonylpyrrolidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (4.57 g, yield: 72.1%).

NMR(CDCl₃) δ: 1.44(9H,s),2.35(3H,s),3.86(1H,m),4.08(1H,m), 4.24(1H,m),5.23(2H,br s),7.53(2H,d,J=8Hz), 8.24(2H,d,J=8Hz) ##STR1267##

A 1N sodium hydroxide aqueous solution (9.7 ml) was added to a solution of the compound obtained by the above reaction (4.56 g, 9.25 mmol) in methanol (100 ml) in a nitrogen stream under cooling with ice. The mixture was stirred at the same temperature for 15 minutes. Triethylamine (2.0 ml, 14.4 mmol) and p-methoxybenzyl chloride (1.82 ml, 13.4 mmol) were added to this reaction solution, and the mixture was stirred at the same temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was extracted with ethyl acetate (300 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 3:1) to obtain (2S,4S)-2-(N-tert-butoxycarbonylpyrrolidin-3-yl)-4-(p-methoxybenzylthio)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (4.97 g, yield: 94.1%).

NMR(CDCl₃) δ: 1.44(9H,s),1.50-1.98(3H,m),2.33(1H,m),2.70(1H,m),3.74 (2H,s),3.80(3H,s),3.96(1H,m),5.20(2H,br s),6.86 (2H,d,J=8Hz),7.24(2H,d,J=8Hz),7.50(2H,d,J=8Hz), 8.24(2H,d,J=8Hz) ##STR1268##

Trifluoroacetic acid (6.7 ml, 10.0 mmol) was added to a solution of the compound obtained by the above reaction (4.97 g, 8.70 mmol) in methylene chloride (50 ml) under cooling with ice, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was extracted with ethyl acetate (150 ml). The organic layer was washed with a saturated sodium hydrogen carbonate aqueous solution and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain (2S,4S)-4-(p-methoxybenzylthio)-N-(p-nitrobenzyloxycarbonyl)-2-(3-pyrrolidinyl)pyrrolidine (4.0 g, yield: 97.6%).

NMR(CDCl₃) δ: 1.62(1H,m),1.75-2.18(2H,m),2.42(1H,m),2.82(1H,m), 3.75(2H,s),3.82(3H,s),4.09(1H,m),5.20 (2H,br s),6.86(2H,d,J=8Hz),7.24(2H,d,J=8Hz),7.50 (2H,d,J=8Hz),8.27(2H,d,J=8Hz) ##STR1269##

A 37% formaldehyde aqueous solution (0.6 ml, 7.5 mmol) and sodium cyanoborohydride (300 mg, 4.77 mmol) were added to a solution of the compound obtained by the above reaction (1.4 g, 2.97 mmol) in acetonitrile (15 ml) at room temperature, and the mixture was stirred at the same temperature for 15 minutes. This reaction mixture was neutralized with acetic acid and then further stirred at room temperature for 1 hour. The reaction mixture was extracted with ethyl acetate (50 ml). The organic layer was washed with a 1N sodium hydroxide aqueous solution, water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate-methanol 1:1) to obtain (2S,4S)-4-(p-methoxybenzylthio)-2-(N-methylpyrrolidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (617 mg, yield: 42.8%).

NMR(CDCl₃) δ: 1.42-2.04(3H,m),3.74(2H,s),3.82(3H,s),3.98(1H,m), 5.20(2H,br s),6.86(2H,d,J=8Hz),7.24(2H,d,J=8Hz), 7.47(2H,br d,J=8Hz),8.25(2H,d,J=8Hz) ##STR1270##

Anisole (0.34 ml, 3.13 mmol) and trifluoromethane-sulfonic acid (0.27 ml, 3.05 mmol) were added to a solution of the compound obtained by the above reaction (617 mg, 1.27 mmol) in trifluoroacetic acid (2.26 ml) at 0° C., and the mixture was stirred at the same temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the residue was repeatedly subjected to decantation with dry diethyl ether to obtain the above identified compound (650 mg, yield: 99%).

REFERENCE EXAMPLE 7 (2S,4S)-4-mercapto-2-(N-methyl-2-azetidinon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine ##STR1271##

A 5.3M methylamine-ethanol solution (5 ml, 26.6 mmol) was added to 3-[(2S,4R)-4-tert-butyldimethylsiloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-2-yl]acrylic acid ethyl ester (2.94 g, 6.15 mmol, compound of Reference Example 1-6 of Japanese Patent Application No. 342,948/1989) at room temperature, and the mixture was left to stand at the same temperature for 2 days. The reaction mixture was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate) to obtain 3-methylamino-3-[(2S,4R)-4-tert-butyldimethylsiloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-2-yl]propionic acid ethyl ester (1.67 g, yield: 53.3%).

NMR(CDCl₃) δ: 0.04(3H,s),0.06(3H,s),0.84(9H,s),1.26(3H,t,J=7Hz), 1.80-2.16(2H,m),2.24-2.50(6H,m),3.40(2H,m), 3.65(1H,m),4.16(2H,m),4.40(1H,m),5.30(2H,m), 7.56(2H,m),8.24(2H,d,J=8Hz) ##STR1272##

The same procedure as in Reference Example 3-2 was carried out by using the compound obtained by the above reaction (1.67 g, 3.28 mmol) to obtain (2S,4R)-4-tert-butyldimethylsiloxy-2-(N-methyl-2-azetidinon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (700 mg, yield: 46.1%).

NMR(CDCl₃) δ: 0.04(3H,s),0.06(3H,s),0.84(9H,s),1.72-2.00(2H,m), 2.55(1H,m),2.80(3H,s),2.94(1H,m),3.36(1H,m), 3.70(1H,m),4.10(1H,m),4.40(2H,m),5.16-5.22(2H,m), 7.67(2H,d,J=8Hz),8.24(2H,d,J=8Hz) ##STR1273##

46% hydrofluoric acid (1 ml) was added to a solution of the compound obtained by the above reaction (700 mg, 1.51 mmol) in acetonitrile (10 ml) at room temperature, and the mixture was stirred at the same temperature for 1 hour. Then, the reaction solution was extracted with ethyl acetate (70 ml). The organic layer was washed sequentially with a 5% sodium hydrogen carbonate aqueous solution, water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 2% methanol-chloroform) to obtain (2S,4R)-4-hydroxy-2-(N-methyl-2-azetidinon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (316 mg, yield: 59.9%).

NMR(CDCl₃) δ: 1.70-2.18(2H,m),2.52(1H,m),2.74(3H,s),2.88(1H,m), 3.60-3.88(2H,m),4.08(1H,m),4.40(2H,m),5.22(2H, br s), 7.50(2H,d,J=8Hz),8.16(2H,d,J=8Hz) ##STR1274##

The same procedure as in Reference Example 3-5 was carried out by using the compound obtained by the above reaction (315 mg, 0.90 mmol) to obtain (2S,4S)-4-acetylthio-2-(N-methyl-2-azetidinon-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (348 mg, yield: 94.7%).

IR(KBr)cm⁻¹ : 1760, 1700, 1520, 1340

NMR(CDCl₃) δ: 1.75(1H,m),2.37(3H,s),2.79(3H,s),3.88(1H,m), 3.98-4.20(3H,m),5.28(2H,br s),7.55(2H,d,J=8Hz), 8.25(2H,d,J=8Hz) ##STR1275##

The same procedure as in Reference Example 1-9 was carried out by using the compound obtained by the above reaction (348 mg, 0.86 mmol) to obtain the above identified compound (298 mg, yield: 95.5%).

REFERENCE EXAMPLE 8 (2S,4S)-4-mercapto-2-(N-methyl-2,5-dioxopyrrolidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomers A and B ##STR1276##

The same procedure as in Reference Example 1-5 was carried out by using 3-[(2S,4R)-4-tert-buthyldimethylsiloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-2-yl]acrylic acid ethyl ester (3.8 g, 7.95 mmol, compound of Reference Example 1-6 of Japanese Patent Application No. 342,948/1989) to obtain 3-(2S,4R)-4-tert-butyldimethylsiloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-2-yl]-4-nitrobutyric acid ethyl ester (4.0 g, yield: 93.3%).

NMR(CDCl₃) δ: 0.04(3H,s),0.06(3H,s),0.82(9H,s),1.26(3H,m), 1.80(1H,m),2.03(1H,m),2.22-2.54(3H,m),3.27(2H,m), 4.15(3H,m),4.32-4.70(2H,m),5.24(2H,br s),7.54(2H,m), 8.25(2H,d,J=8Hz) ##STR1277##

Sodium nitrite (2.56 g, 37.1 mmol) was added to a solution of the compound obtained by the above reaction (4.0 g, 7.42 mmol) in dimethyl sulfoxide (40 ml) in a nitrogen stream at room temperature. Then, butyl nitrite (1.74 ml, 14.9 mmol) was dropwise added thereto, and the mixture was stirred overnight at the same temperature. The reaction solution was extracted with ethyl acetate (150 ml). The organic layer was washed three times with water and once with a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was dissolved in tetrahydrofuran (50 ml). Then, triethylamine (0.97 ml, 6.95 mmol) and isobutyl chloroformate (0.90 ml, 6.94 mmol) were dropwise added thereto in a nitrogen stream at -20° C., and the mixture was stirred at the same temperature for 30 minutes. Then, 40% methylamine (1 ml) was added thereto. The reaction mixture was stirred for 30 minutes under cooling with ice and then extracted with ethyl acetate (100 ml). The organic layer was washed seqnentially with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate) to obtain (2S,4R)-4-tert-butyldimethylsiloxy-2-(2-ethoxycarbonyl-1-methylcarbamoylethyl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (1.8 g, yield: 45.6%).

NMR(CDCl₃) δ: 0.04(3H,s),0.05(3H,s),0.83(9H,s),1.24(3H,t,J=7Hz, 3.34(1H,m),3.56(1H,m),4.12(2H,m),4.32(1H,m), 5.28(2H,m),7.52(2H,m),8.25(2H,m) ##STR1278##

A 1N sodium hydroxide aqueous solution (3.7 ml) was dropwise added to a solution of the compound obtained by the above reaction (1.8 g, 3.35 mmol) in ethanol (30 ml) at room temperature, and the mixture was stirred overnight at the same temperature. 1N hydrochloric acid (3.7 ml) was added to the reaction solution, and the mixture was concentrated under reduced pressure. Then, the residue was extracted with ethyl acetate (70 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was dissolved in acetic anhydride (15 ml), and sodium acetate (1.38 g, 16.8 mmol) was added thereto. The mixture was stirred at 100° C. for 1.5 hours. The reaction mixture was concentrated under reduced pressure, and then the residue was extracted with ethyl acetate (70 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 3:1) to obtain (2S,4R)-4-tert-butyldimethylsiloxy-2-(N-methyl-2,5-dioxopyrrolidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (1.28 g, yield: 77.8%).

NMR(CDCl₃) δ: 0.04(3H,s),0.06(3H,s),0.84(9H,s),1.71(1H,m), 2.96(3H,s),3.16-3.87(3H,m),4.37(1H,m),4.54(1H,m), 5.22(2H,m),7.50(2H,m),8.20(2H,m) ##STR1279##

The same procedures as in Reference Example 7-3 and 7-4 were carried out by using the compound obtained by the above reaction (1.28 g, 2.61 mmol) to obtain (2S,4S)-4-acetylthio-2-(N-methyl-2,5-dioxopyrrolidin-3-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer A (406 mg, yield: 35.8%) and diastereomer B-(170 mg, yield: 15%), respectively.

Diastereomer A

NMR(CDCl₃) δ: 1.60(1H,m),2.34(3H,s),2.97(3H,s),3.60-3.98(2H,m), 4.22(1H,m),4.47(1H,m),5.24(2H,br s),7.56(2H,d,J=8Hz), 8.24(2H,d,J=8Hz)

Diastereomer B

IR(KBr)cm⁻¹ : 1700, 1520, 1440, 1350

NMR(CDCl₃) δ: 1.72(1H,m),2.36(3H,s),2.97(3H,br s),3.25(2H,m), 3.88(1H,m),4.18(1H,m),4.42(1H,m),5.07-5.30(2H,m), 7.51(2H,d,J=8Hz),8.24(2H,d,J=8Hz) ##STR1280##

The same procedure as in Reference Example 1-9 was carried out by using the diastereomer A obtained by the above reaction (406 mg, 0.93 mmol) to obtain diastereomer A of the above identified compound (330 mg, yield: 90%). ##STR1281##

The same procedure as in Reference Example 1-9 was carried out by using the diastereomer B obtained in Reference Example 8-4 (170 mg, 0.39 mmol) to obtain diastereomer B of the above identified compound (145 mg, yield: 94.6%).

REFERENCE EXAMPLE 9 (2S,4S)-N-allyloxycarbonyl-2-(2,5-dioxopyrrolidin-3-yl)-4-mercaptopyrrolidine ##STR1282##

The same procedure as in Reference Exmample 1-5 was carried out by using 3-[(2S,4S)-N-allyloxycarbonyl-4-tritylthiopyrrolidin-2-yl]acrylic acid ethyl ester (5.23 g, 9.9 mmol) which was obtained by a reaction similar to Reference Example 1-4, to obtain 3-[(2S,4S)-N-allyloxycarbonyl- 4-tritylthiopyrrolidin-2-yl]-4-nitrobutyric acid ethyl ester (4.74 g, yield: 81.2%).

NMR(CDCl₃) δ: 1.24(3H,t,J=8Hz),3.92(1H,m),4.12(2H,q,J=8Hz), 4.47(3H,m),5.26(2H,m),5.86(1H,m),7.16-7.74(15H,m) ##STR1283##

Sodium nitrite (2.76 g, 40 mmol) was added to a solution of the compound obtained by the above reaction (4.7 g, 8.0 mmol) in dimethyl sulfoxide (35 ml) in a nitrogen stream at room temperature. Then, butyl nitrite (1.87 ml, 16 mmol) was dropwise added thereto, and the mixture was stirred overnight at the same temperature. The reaction solution was extracted with ethyl acetate (150 ml). The organic layer was washed three times with water and once with a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 1% methanol-chloroform) to obtain (2S,4S)-N-allyloxycarbonyl-2-(1-carboxy-2-ethoxycarbonylethyl)-4-tritylthiopyrrolidine (1.9 g, yield: 41.5%).

NMR(CDCl₃) δ: 1.23(3H,m), 3.90-4.24(3H,m),4.48(2H,m), 5.18-5.32(2H,m),5.86(1H,m), 7.16-7.60(15H,m) ##STR1284##

Triethylamine(0.56 ml, 4.0 mmol) and isobutyl chloroformate (0.52 ml, 4.0 mmol) were dropwise added to a solution of the compound obtained by the above reaction (1.9 g, 3.3 mmol) in tetrahydrofuran (30 ml) in a nitrogen stream at -20° C. and the mixture was stirred at the same temperature for 30 minutes. Then, concentrated aqueous ammonia (0.7 ml, 10.5 mmol) was added thereto, and the mixture was stirred further for 30 minutes under cooling with ice. The reaction solution was extracted with ethyl acetate (100 ml). The organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 5:1) to obtain a polar compound (744 mg, yield: 39.2%) and a less polar compound (711 mg, yield: 37.5%) of (2S,4S)-N-allyloxycarbonyl-2-(1-carbamoyl-2-ethoxycarbonylethyl)-4-tritylthiopyrrolidine.

Polar compound

NMR(CDCl₃) δ: 1.23(3H,t,J=7Hz),1.98(2H,m),3.85(1H,m),4.15 (2H,q,J=7Hz),4.50(2H,m),5.24(2H,m),5.85(1H,m), 7.15-7.60(15H,m)

Less polar compound

NMR(CDCl₃) δ: 1.23(3H,t,J=7Hz),1.95(2H,m),2.26(1H,m),3.76(1H,m), 4.12(2H,q,J=7Hz),4.50(2H,m),5.24(2H,m),5.84(1H,m), 7.14-7.57(15H,m) ##STR1285##

Sodium hydride (57 mg, 1.43 mmol) was added to, a solution of the polar compound obtained by the above reaction (744 mg, 1.3 mmol) in tetrahydrofuran (37 ml) under cooling with ice, and the mixture was stirred at the same temperature for 15 minutes. To the reaction solution, water (1 ml) and then 1N hydrochloric acid (1.43 ml) were added, and the mixture was extracted with ethyl acetate (70 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 3:1) to obtain (2S,4S)-N-allyloxycarbonyl-2-(2,5-dioxopyrrolidin-3-yl)-4-tritylthiopyrrolidine (630 mg, yield: 92.1%).

NMR(CDCl₃) δ: 1.40(1H,m),2.14(1H,m),2.15-2.95(4H,m),3.88(1H,m), 4.10(1H,m),4.47(2H,m),5.13-5.36(2H,m),5.84(1H,m), 7.10-7.60(15H,m) ##STR1286##

The same procedure as in Reference Example 22-2 was carried out by using the compound obtained by the above reaction (630 mg, 1.2 mmol) to obtain the above identified compound (219 mg, yield: 64.4%).

NMR(CDCl₃ -CD₃ OD) δ: 2.38-2.78(4H,m),2.95-3.26(2H,m),3.80-4.22(1H,m), 4.34(1H,m),4.56(2H,m),5.16-5.38(2H,m),5.90(1H,m)

REFERENCE EXAMPLE 10 (2S,4S)-N-allyloxycarbonyl-4-mercapto-2-1-allyloxycarbonyl-3-pyrazolidinon-5-yl)pyrrolidine ##STR1287##

Hydrazine monohydrate (0.61 ml, 12.6 mmol) was dropwise added to a solution of 3-[(2S,4S)-N-allyloxycarbonyl-4-tritylthiopyrrolidin-2-yl]acrylic acid ethyl ester (1.53 g, 2.9 mmol) in ethanol (7.5 ml) under cooling with ice, and the mixture was stirred at the same temperature for 1.5 hours. The reaction solution was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 2% methanol-chloroform) to obtain (2S,4S)-N-allyloxycarbonyl-2-(3-pyrazolidinon-5-yl)-4-tritylthiopyrrolidine (830 mg, yield: 51.1%). This compound was immediately dissolved in methylene chloride (10 ml). Then, triethylamine (0.24 ml, 1.7 mmol) and allyl chloroformate (0.18 ml 1.7 mmol) were dropwise added thereto under cooling with ice, and the mixture was stirred at the same temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the residue was extracted with ethyl acetate (50 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 1% methanol-chloroform) to obtain (2S,4S)-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-3-pyrazolidinon-5-yl)-4-tritylthiopyrrolidine (592 mg, yield: 62%).

NMR(CDCl₃) δ: 1.46-1.88(2H,m),1.94-2.24(2H,m),2.56-3.00(4H,m), 3.70(1H,m),4.34-4.50(4H,m),5.12-5.40(4H,m),5.75-6.00 (2H,m),7.15-8.60(15H,m) ##STR1288##

The same procedure as in Reference Example 22-2 was carried out by using the compound obtained by the above reaction (592 mg, 0.92 mmol) to obtain (2S,4S)-N-allyloxycarbonyl-4-mercapto-2-(1-allyloxycarbonyl-3-pyrazolidinon-5-yl)pyrrolidine (324 mg, yield: 87.8%).

NMR(CDCl₃) δ: 1.60-1.95(2H,m),2.30(1H,dd,J=17,3Hz),2.56(1H,m), 2.88-3.30(3H,m),3.88-4.24(2H,m),4.52-4.98(4H,m), 5.00-5.45(4H,m),5.82-6.08(2H,m)

REFERENCE EXAMPLE 11 (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidony-3-yl)pyrrolidine ##STR1289##

The same procedure as in Reference Example 9-2 was carried out by using 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-4-nitrobutyric acid ethyl ester obtained in Reference Example 1-5 (5.9 g, 12.8 mmol) to obtain (2S,4R)-2-(2-ethoxycarbonyl-1-carboxyethyl)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonylpyrrolidine (3.57 g, yield: 62.5%).

NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,s),1.26(3H,t,J=8Hz),1.48(9H,s), 2.01(1H,m),2.34(1H,m),2.38(1H,m),3.25 (1H,dd,J=12,4Hz),3.38-3.90(3H,m),4.16(2H,q,J=8Hz), 4.25-4.55(2H,m) ##STR1290##

The same procedure as in Reference Example 9-3 was carried out by using the compound obtained by the above reaction (3.57 g, 8.0 mmol) and benzylamine (1.55 ml, 14.2 mmol) to obtain (2S,4R)-2-(1-benzylcarbamoyl-2-ethoxycarbonylethyl)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonylpyrrolidine (3.7 g, yield: 86.3%). NMR(CDCl₃) δ: 0.04(6H,s),0.85(9H,s),1.24(3H,t,J=8Hz),1.33-1.55 (9H,m),1.90(1H,m),2.30(1H,m),2.82(1H,m),3.22(1H,m), 3.35-3.82(2H,m),4.00-4.65(5H,m),7.30(5H,m) ##STR1291##

A 1N sodium hydroxide aqueous solution (3.9 ml) was added to a solution of the compound obtained by the above reaction (1.89 g, 3.5 mmol) in ethanol (30 ml) at room temperature, and the mixture was stirred overnight at the same temperature. 1N hydrochloric acid (3.9 ml) was added to the reaction solution, and the mixture was concentrated under reduced pressure to obtain (2S,4R)-2-(1-benzylcarbamoyl-2-carboxyethyl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidine (1.79 g, yield: 100%). A borane-dimethyl sulfide complex (0.72 ml, 7.2 mmol) was dropwise added to a solution of this compound in tetrahydrofuran (35 ml) in a nitrogen stream at room temperature, and the mixture was stirred at the same temperature for 1 hour. Methanol (5 ml) was added to the reaction solution, and the mixture was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 1:1) to obtain 3-benzylcarbamoyl-3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-1-propanol (1.27 g, yield: 72.9%).

NMR(CDCl₃) δ: 0.04(6H,s),0.86(9H,s),1.24-1.60(1H,m),1.73-2.32 (2H,m),3.32-3.73(3H,m),4.12-4.34(2H,m),4.40(1H,m), 7.30 (5H,m) ##STR1292##

A solution of the compound obtained by the above reaction (1.27 g, 2.6 mmol) in hexamethylphosphoric triamide (17 ml) was dropwise added to a solution of potassium tert-butoxide (640 mg, 5.7 mmol) in tetrahydrofuran (17 ml) in a nitrogen stream at 0° C., and the mixture was stirred at the same temperature for 1 hour. A solution of p-toluenesulfonyl chloride (520 mg, 2.7 mmol) in tetrahydrofuran (3 ml) was dropwise added to the reaction solution, and the mixture was stirred at 0° C. for 1 hour and at 50° C. for 2 hours. The reaction mixture was cooled with ice, and a saturated ammonium chloride aqueous solution (5 ml) was added thereto. The mixture was extracted with ethyl acetate (100 ml). The organic layer was washed three times with water and once with a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C- 300, hexane-ethyl acetate 10:1) to obtain (2S,4R)-2-(N-benzyl-2-pyrrolidon-3-yl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidine (343 mg, yield: 39.8%).

NMR(CDCl₃) δ: 0.06(6H,s),0.87(9H,s),1.46(9H,s),1.90-2.30(3H,m), 3.00-3.60(5H,m),4.18-4.64(5H,m),7.18-7.42(5H,m) ##STR1293##

The same procedure as in Reference Example 3-3 was carried out by using the compound obtained by the above reaction (760 mg, 1.6 mmol) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-pyrrolidon-3-yl)pyrrolidine (476 mg, yield: 77.2%).

NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,s),1.46(9H,s),1.82(1H,m),2.04 (2H,m),3.20-3.70(5H,m),4.32(1H,m),4.42(1H,m) ##STR1294##

The same procedure as in Reference Example 3-4 was carried out by using the compound obtained by the above reaction (476 mg, 1.24 mmol) to obtain (2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-3-yl)pyrrolidine (342 mg, yield: 79.1%).

IR(KBr)cm⁻¹ : 1700, 1520, 1350

NMR(CDCl₃) δ: 1.70-2.28(4H,m),3.18-3.50(3H,m),3.52-3.88(2H,m), 4.28-4.68(2H,m),5.22(2H,m),7.53(2H,d,J=8Hz), 8.20(2H,d,J=8Hz) ##STR1295##

The same procedure as in Reference Example 3-5 was carried out by using the compound obtained by the above reaction (342 mg, 0.98 mmol) to obtain (2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-2-yl)pyrrolidine (271 mg, yield: 68.2%).

IR(KBr)cm⁻¹ : 1700, 1530, 1520

NMR(CDCl₃) δ: 1.62-2.18(3H,m),2.35(3H,s),3.02-3.70(5H,m), 3.83(1H,m),4.25(1H,m),4.44(1H,m),5.28(2H,br s), 7.56(2H,d,J=8Hz) ,8.25(2H,d,J=8Hz) ##STR1296##

The same procedure as in Reference Example 1-9 was carried out by using the compound obtained by the above reaction (271 mg, 0.67 mmol) to obtain the above identified compound (220 mg, yield: 90.5%).

REFERENCE EXAMPLE 12 (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomers A and B ##STR1297##

To the compound of Reference Example 1-6 (10.82 g, 22.32 mmol), n-hexane (75 ml) was added, and the mixture was heated to 40° C. for dissolution and then filtered. The filtrate was left to stand overnight at room temperature. The precipitate was collected by filtration and dried to obtain a polar compound (diastereomer A; 3.74 g, yield: 34.6%) of the above identified compound. The filtrate was concentrated, and n-hexane (70 ml) was added to the residue. Then, the above operation was repeated to obtain a low polar compound (diastereomer B; 1.87 g, yield: 17.3%) of the above identified compound.

Diastereomer A (Polar compound)

mp: 92°-95° C.

[α]_(D) ²⁰ : -50.8° (C=1.031), MeOH)

IR(KBr)cm⁻¹ : 1685, 1395, 1250, 1175

NMR(CDCl₃) δ: 0.06(6H,s), 0.86(9H,s),1.46(9H,s),1.72-1.97(2H,m), 2.2(1H,dd,J=18,8Hz),2.41(1H,dd,J=18,8Hz),2.9-3.8 (5H,m),4.15(1H,br),4.32(1H,br),6.06(1H,br)

Diastereomer B (Less polar compound)

mp: 97°-100° C.

[α]_(D) ²⁰ : 53.9° (C=0.978, MeOH)

IR(KBr)cm⁻¹ : 1685, 1665, 1385, 1255, 1160

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.46(9H,s),1.7-1.95(2H,m),2.04 (1H,dd,J=16,8Hz),2.32(1H,dd,J=16,8Hz),2.9-3.6 (5H,m),4.1(1H,br),4.32(1H,br),6.0(1H,br)

HPLC:

Column: Daicel CHIRALCEL OD

Eluent: Hexane:isopropanol=9:1

Flow rate: 0.5 ml/min

Temperature: 38° C.

Detector: 210 nm

Retention time:

Diastereomer A:19.8 min

Diastereomer B:16.3 min

REFERENCE EXAMPLE 13 (2S,4S)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer I ##STR1298##

The same procedure as in Reference Example 6-2 was carried out by using the (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer A (polar compound; 8.0 g, 20.8 mmol) obtained in Reference Example 12 to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonyl-2-pyrrolidon-4-yl)pyrrolidine diastereomer A (9.5 g, yield: 94.2%).

NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,s),1.46(9H,s),1.53(9H,s),1.70 (1H,m),1.97(1H,m),2.16-2.90(3H,m),3.24 (1H,dd,J=12,4Hz),3.36-3.70(2H,m),3.76 (1H,dd,J=10,8Hz),4.12(1H,m),4.33(1H,m) ##STR1299##

A 1M vinyl magnesium bromide-tetrahydrofuran solution (23 ml) was dropwise added to a solution of the compound obtained by the above reaction (9.3 g, 19.2 mmol) in tetrahydrofuran (120 ml) in a nitrogen stream at -40° C., and the mixture was stirred at the same temperature for 2 hours. A 50% acetic acid-methanol solution (10 ml) was dropwise added to the reaction solution, and the mixture was extracted with ethyl acetate (200 ml). The organic layer was washed sequantially with a saturated sodium hydrogen carbonate aqueous solution, water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 10:1) to obtain 5-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-6-tert-butoxycarbonylamino-1-hexen-3-one diastereomer A (4.1 g, yield: 41.7%).

NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,s),1.42(9H,s),1.46(9H,s), 4.04(1H,m),4.24(1H,m),5.80(1H,br d,J=10Hz),6.22 (1H,d,J=17Hz),6.38(1H,dd,J=17,10Hz) ##STR1300##

A solution of cerium chloride hexahydrate (2.84 g, 8.0 mmol) in methanol (30 ml) was added to the compound obtained by the above reaction (4.1 g, 8.0 mmol). Then, sodium borohydride (310 mg, 8.2 mmol) was added thereto at -15° C., and the mixture was stirred at the same temperature for 15 minutes. Water (10 ml) was added to the reaction mixture, and the mixture was concentrated under reduced pressure. The residue was extracted with ethyl acetate (100 ml). The organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 8:1) to obtain 5-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-6-tert-butoxycarbonylamino-1-hexen-3-ol (3.53 g, yield: 85.8%).

NMR(CDCl₃) δ: 0.04(6H,s),0.86(9H,s),1.43(9H,s),1.46(9H,s),4.30 (2H,m),5.12(1H,br d,J=10Hz),5.28(1H,br d,J=16Hz), 5.90(1H,m) ##STR1301##

A solution of the compound obtained by the above reaction (3.53 g, 6.9 mmol) in tetrahydrofuran (15 ml) was dropwise added to a solution of potassium tert-butoxide (1.7 g, 15.1 mmol) in tetrahydrofuran (50 ml) in a nitrogen stream at -10° C., and the mixture was stirred at the same temperature for 30 minutes. Then, a solution of p-toluenesulfonyl chloride (1.44 g, 7.5 mmol) in tetrahydrofuran (10 ml) was dropwise added thereto. The reaction solution was stirred at the same temperature for 1 hours. Then, a saturated ammonium chloride aqueous solution (10 ml) was added thereto, and the mixture was extracted with ethyl acetate (100 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 10:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonyl-2-vinylpyrrolidin-4-yl)pyrrolidine (2.09 g, yield: 61.5%).

NMR(CDCl₃) δ: 0.04(6H,s),0.86(9H,s),1.42(9H,s),1.45(9H,s),2.98 (1H,m),3.14(1H,dd,J=12,4Hz),3.32-3.72(2H,m),3.92-4.20 (2H,m),4.34(1H,m),4.96-5.20(2H,m),5.74(1H,m) ##STR1302##

Water (13 ml) and sodium periodate (3.70 g, 17.3 mmol) were added to a solution of the compound obtained by the above reaction (2.09 g, 4.2 mmol) in a mixture of carbon tetrachloride (10 ml) and acetonitrile (10 ml), and the mixture was vigorously stirred. Ruthenium chloride hydrate (20 mg, 0.096 mmol) was added to this reaction mixture, and the mixture was vigorously stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, and the residue was extracted with ethyl acetate (100 ml). The organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 1% methanol-chloroform) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonyl-2-carboxypyrrolidin-4-yl)pyrrolidine (1.07 g, yield: 50.7%).

NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,s),1.46(18H,br s),3.10(1H,m), 3.24(1H,m),3.40-3.74(2H,m),3.96-4.40(3H,m) ##STR1303##

The same procedure as in Reference Example 9-3 was conducted by using the compound obtained by the above reaction (1.07 g, 2.1 mmol) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonyl-2-carbamoylpyrrolidin-4-yl)pyrrolidine (867 mg, yield: 81.2%).

NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,s),1.46(18H,s), 3.06(1H,br t,J=12Hz),3.23(1H,dd,J=12,4Hz), 3.36-3.82(2H,m),3.98-4.40(3H,m) ##STR1304##

The same procedure as in Reference Example 5-3 was carried out by using the compound obtained by the above reaction (861 mg, 1.68 mmol) to obtain (2S,4R)-2-2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer I (polar compound, 621 mg, yield: 66.4%) and diastereomer II (less polar compound, 216 mg, yield: 23.1%).

Polar compound

NMR(CDCl₃) δ: 3.78(2H,m),4.26(2H,m),4.50(1H,m),5.26(4H,br s),7.54 (4H,d,J=8Hz),8.25(4H,d,J=8Hz)

Less polar compound

NMR(CDCl₃) δ: 3.70(2H,m),4.00-4.53(3H,m),5.24(4H,br s),7.54 (4H,d,J=8Hz),8.20(4H,d,J=8Hz) ##STR1305##

The same procedure as in Reference Example 5-4 was carried out by using the polar compound obtained by the above reaction (621 mg, 1.1 mmol) to obtain (2S,4R)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer I (670 mg, yield: 94.6%).

NMR(DMSO-d₆) δ: 3.25(3H,s),3.48-3.70(2H,m),3.92-4.22(3H,m),5.04-5.34 (5H,m),7.64(4H,d,J=8Hz),8.24(4H,m) ##STR1306##

Sodium iodide (174 mg, 1.16 mmol) and potassium thioacetate (240 mg, 2.1 mmol) were added to a solution of the compound obtained by the above reaction (670 mg, 1.06 mmol) in N,N-dimethylformamide (10 ml) in a nitrogen stream, and the mixture was stirred overnight at a temperature of from 60° to 70° C. The reaction solution was extracted with ethyl acetate (70 ml). The organic layer was washed three times with water and once with a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 1% methanol-chloroform) to obtain (2S,4S)-4-acetylthio-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer I (479 mg, yield: 73.8%).

NMR(CDCl₃) δ: 1.74(1H,m),2.00(1H,m),2.35(3H,s),2.40-2.90(3H,m), 5.24(4H,br s),7.54(4H,br d,J=8Hz),8.22(4H,br d,j=8Hz) ##STR1307##

The same procedure as in Reference Example 1-9 was carried out by using the compound obtained by the above reaction (479 mg, 0.78 mmol) to obtain (2S,4S)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer I (280 mg, yield: 62.7%).

REFERENCE EXAMPLE 14 (2S,4S)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer II ##STR1308##

The same procedure as in Reference Example 5-4 was carried out by using the (2S,4R)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidin (less polar compound, 216 mg, 0.39 mmol) obtained in Reference Example 13-7, to obtain (2S,4R)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer I: (221 mg, yield: 89.7%).

NMR(CDCl₃) δ: 3.04(3H,s),3.58(2H,m),4.08-4.32(2H,m), 4.42(1H,m),5.10-5.38(5H,m),5.50-5.68(1H,m) ##STR1309##

The same procedure as in Reference Example 13-9 was carried out by using the compound obtained by the above reaction (221 mg, 0.35 mmol) to obtain (2S,4S)-4-acetylthio-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer II (180 mg, yield: 84.0%).

NMR(CDCl₃) δ: 1.66(1H,m),1.88-2.28(2H,m),2.36(3H,s),2.54(1H,m), 2.82(1H,m),3.06-3.35(2H,m),3.63(1H,m), 3.86(1H,m), 4.08(1H,m),4.27(1H,m),4.47(1H,m),5.62(4H,m), 7.54(4H,d,J=8Hz),8.24(4H,d,J=8Hz) ##STR1310##

The same procedure as in Reference Example 1-9 was carried out by using the compound obtained by the above reaction (180 mg, 0.29 mmol) to obtain (2S,4S)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer II (160 mg, yield: 95%).

REFERENCE EXAMPLE 15 (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]pyrrolidine diastereomer A ##STR1311##

The same procedure as in Reference Example 5-1 was carried out by using the (2S,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonyl-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer A (4.8 g, 12.0 mmol) obtained in Reference Example 12 to obtain crude (2S,4R)-4-tert-butyldimethylsiloxycarbonyl-2-(N-tert-butoxycarbonyl-2-pyrrolidon-4-yl)pyrrolidine diastereomer A (6.8 g).

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.48(9H,s),1.52(9H,s),1.60 (1H,m),1.90(1H,m),2.10-2.58(2H,m),3.22(1H,m), 3.42-3.88(3H,m),4.08(1H,m),4.30(1H,m) ##STR1312##

The same procedure as in Reference Example 5-2 was carried out by using the compound obtained by the above reaction (6.8 g) to obtain a yellow oily substance (5.7 g) of (2S,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonyl-2-(N-tert-butoxycarbonylpyrrolidin-4yl)pyrrolidine diastereomer A, which was used for the subsequent reaction without purification.

NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,s),1.44(18H,s),1.56-2.04(4H,m), 3.02-3.70(6H,m),4.00(1H,m),4.34(1H,m) ##STR1313##

The compound obtained by the above reaction (5.7 g) was added to an about 3N hydrogen chloride-methanol solution, and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure to obtain crystals of (2S,4R)-4-hydroxy-2-(3-pyrrolidinyl)pyrrolidine diastereomer A dihydrochloride (2.4 g).

IR(KBr)cm⁻¹ : 3300, 2900, 2700, 1430, 1410, 1060

NMR(D₂ O) δ: 1.92(2H,m),2.16-2.46(2H,m),2.68(1H,m), 3.06(1H,dd,J=12,10Hz),3.34(2H,br d,J=12Hz), 3.42-3.66(3H,m),3.82(1H,m),4.67(1H,m) ##STR1314##

The same procedure as in Reference Example 5-3 was carried out by using the compound obtained by the above reaction (1.1 g) to obtain an oily substance (2.2 g, yield: 88%) of (2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]pyrrolidine diastereomer A. ##STR1315##

The same procedure as in Reference Example 5-4 was carried out by using the compound obtained by the above reaction (2.2 g) to obtain a foamy substance (2.6 g) of (2S,4R)-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]pyrrolidine diastereomer A.

NMR(CDCl₃) δ: 2.02(2H,m),2.50(2H,m),3.04(3H,s),3.14-3.82(6H,m), 4.20(2H,m),5.24(4H,br s),7.53(4H,d,J=8Hz), 8.24(4H,d,J=8Hz) ##STR1316##

The same procedure as in Reference Example 5-5 was carried out by using the compound obtained by the above reaction (2.6 g) to obtain (2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]pyrrolidine (1.7 g, yield from the first step: 73%).

NMR(CDCl₃) δ: 1.54-2.10(4H,m),2.36(3H,s),2.40-2.82(2H,m), 3.02-3.70(6H,m),3.88(1H,m),4.00-4.32(2H,m), 5.24(4H,br s),7.52(4H,d,J=8Hz),8.24(4H,d,J=8Hz) ##STR1317##

The same procedure as in Reference Example 1-9 was carried out by using the compound obtained by the above reaction (1.7 g) to obtain the above identified compound (1.59 g).

REFERENCE EXAMPLE 16 (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]pyrrolidine diastereomer B ##STR1318##

The same procedure as in Reference Example 5-1 was carried out by using the (2S,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonyl-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer B (5.1 g, 13.3 mmol) obtained in Reference Example 12, to obtain crude (2S,4R)-4-tert-butyldimethylsiloxycarbonyl-2-(N-tert-butoxycarbonyl-2-pyrrolidon-4-yl)pyrrolidine diastereomer B (7.5 g). ##STR1319##

The same procedure as in Reference Example 5-2 was carried out by using the compound obtained by the above reaction (7.5 g) to obtain a yellow oily substance (6.9 g) of (2S,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonyl-2-(N-tert-butoxycarbonylpyrrolidin-4-yl)pyrrolidine diastereomer B, which was used for the subsequent reaction without purification. ##STR1320##

The compound obtained by the above reaction (6.9 g) was added to an about 3N hydrogen chloride-methanol solution, and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure to obtain crystals of (2S,4R)-4-hydroxy-2-(3-pyrrolidinyl)pyrrolidine diastereomer B dihydrochloride (3.0 g). ##STR1321##

The same procedure as in Reference Example 5-3 was carried out by using the compound obtained by the above reaction (3.0 g) to obtain an oily substnace (6.8 g, yield: 99%) of (2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]pyrrolidine diastereomer B. ##STR1322##

The same procedure as in Reference Example 5-4 was carried out by using the compound obtained by the above reaction (6.8 g) to obtain a foamy substance (7.8 g) of (2S,4R)-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]pyrrolidine diastereomer B. ##STR1323##

The same procedure as in Reference Example 5-5 was carried out by using the compound obtained by the above reaction (3.6 g, 5.8 mmol) to obtain (2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]pyrrolidine diastereomer B (1.3 g, yield: 39%).

NMR(CDCl₃) δ: 1.54-2.10(4H,m),2.36(3H,s),2.40-2.82(2H,m),3.02-3.70 (6H,m),3.88(1H,m),4.00-4.32(2H,m),5.24(4H,br s),7.52 (4H,d,J=8Hz),8.24(4H,d,J=8Hz) ##STR1324##

The same procedure as in Reference Example 1-9 was carried out by using the compound obtained by the above reaction (1.7 g) to obtain the above identified compound (1.6 g).

REFERENCE EXAMPLE 17 (2S,4R)-4-hydroxy-2-(3-pyrrolidinyl)pyrrolidine dihydrochloride ##STR1325##

The same procedure as in Reference Example 15-3 was carried out by using (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonylpyrrolidin-3-yl)pyrrolidine (308 q, 0.654 mol, compound of Reference Example 5-2) to obtain the above identified compound (133 g, yield: 88% ).

IR(KBr)cm⁻¹ : 3390, 3000, 2400, 1600, 1420, 1270, 1065, 980

NMR(D₂ O) δ: 1.76-2.18(2H,m),2.20-2.48(2H,m),2.58-2.98(2H,m), 3.10-3.28(1H,m),3.30-3.50(2H,m),3.50-3.82(3H,m), 3.82-4.04(1H,m),4.70(1H,m)

REFERENCE EXAMPLE 18 (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer A ##STR1326##

The same procedure as in Reference Example 1-7 was carried out by using (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer A (61.38 g, 160 mmol, compound of Reference Example 12) to obtain (2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer A (41.68 g, yield: 74.6%).

IR(KBr)cm⁻¹ : 1695, 1605, 1520, 1430, 1345, 1110

NMR(CDCl₃) δ: 1.6-2.0(2H,m),2.0-2.54(2H,m),3.0-3.56(4H,m), 3.8(1H,m),4.26(1H,m),4.5(1H,m),5.25(2H,s), 5.83(1H,br),7.54(2H,d,J=9Hz),8.22(2H,d,J=9Hz) ##STR1327##

The same procedure as in Reference Example 5-4 was carried out by using the compound obtained by the above reaction (41.6 g, 120 mmol) to obtain (2S,4R)-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer A (47.28 g, yield: 92.3%).

IR(KBr)cm⁻¹ : 1710, 1695, 1605, 1525, 1345, 1170

NMR(CDCl₃) δ: 1.5-2.1(4H,m),2.4-2.8(2H,m),3.04(3H,s),3.1-3.64 (3H,m),4.0-4.5(2H,m),5.2(2H,s),6.32(1H,br), 7.52(2H,d,J=9Hz),8.22(2H,d,J=9Hz) ##STR1328##

The same procedure as in Reference Example 5-5 was carried out by using the compound obtained by the above reaction (13 g, 30 mmol) to obtain (2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer A (4.61 g, yield: 37.7%).

IR(KBr)cm⁻¹ : 1705, 1695, 1515, 1405, 1345, 1110

NMR(CDCl₃) δ: 1.68(2H,m),2.04-2.64(4H,m),2.35(3H,s),3.18(2H,m), 3.42(1H,m),3.84(1H,m),4.04-4.34(2H,m),5.23(2H,s), 5.97(1H,br),7.53(2H,d,J=9Hz),8.25(2H,d,J=9Hz) ##STR1329##

The same procedure as in Reference Example 5-6 was carried out by using the compound obtained by the above reaction (4.61 g, 11 mmol) to obtain the above identified compound (3.92 g, yield: 97.5%).

IR(KBr)cm⁻¹ : 1695, i525, 1400, 1345, 1110

NMR(CDCl₃) δ: 1.42-1.8(2H,m),1.73(1H,d,J=8Hz),2.04-2.6(3H,m), 2.98-3.48(5H,m),4.10(2H,m),5.24(2H,s),6.0(1H,br), 7.52(2H,d,J=9Hz),8.24(2H,d,J=9Hz)

REFERENCE EXAMPLE 19 (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer B ##STR1330##

(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer B (62.48 g, 163 mmol, compound of Reference Example 12) was dissolved in methanol (600 ml), and the solution was cooled to 0° C. Then, a 2.5N hydrochloric acid/methanol solution (230 ml, 570 mmol) was added thereto, and the mixture was stirred at room temperature for 3 hours. The precipitate was collected by filtration and dried to obtain white powder (30.01 g, yield: 89.1%) of (2S,4R)-4-hydroxy-2-(2-pyrrolidon-4-yl)pyrrolidine hydrochloride. This white powder (10.53 g, 50 mmol) was dissolved in a solvent mixture of dioxane (50 ml) and water (50 ml), and the pH was adjusted to 8 with triethyl amine. Then, 4,6-dimethyl-2-(p-nitrobenzyloxycarbonylthio)pyrimidine (17.55 g, 55 mmol) was added thereto, and the mixture was reacted for 6 hours while maintaining the pH at a level of 8. The reaction solution was concentrated, and tetrahydrofuran (100 ml) was added thereto. Insoluble matters were filtered off, and the filtrate was concentrated. The residue was subjected to column chromatography (Wakogel™ C-300, chloroform-methanol 40:1) to obtain (2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4yl)pyrrolidine diastereomer B (17.48 g, yield: 98.2%.

IR(KBr)cm⁻¹ : 1690, 1680, 1525, 1345, 1110, 1095

NMR(CDCl₃) δ: 1.5-1.96(2H,m),1.98-2.46(2H,m),3.0-3.56(4H,m), 3.7-3.9(1H,m),4.25(1H,m),4.5(1H,br),5.26(2H,s), 6.02(1H,br),7.54(2H,d,J=9Hz),8.25(2H,d,J=9Hz) ##STR1331##

The same procedure as in Reference Example 5-4 was carried out by using the compound obtained by the above reaction (17.46 g, 50 mmol) to obtain (2S,4R)-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer B (17.46 g, yield: 81.8%).

IR(KBr)cm⁻¹ : 1705, 1690, 1520, 1345, 1170, 905

NMR(CDCl₃) δ: 1.74(2H,br),1.9-2.6(4H,m),3.04(3H,s),3.2-3.64(3H,m), 4.1-4.36(2H,m),5.28(2H,s),5.92(1H,br),7.54 (2H,d,J=9Hz),8.24(2H,d,J=9Hz) ##STR1332##

The same procedure as in Reference Example 5-5 was carried out by using the compound obtained by the above reaction (17.4 g, 41 mmol) to obtain (2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer B (14.29 g, yield: 85.6%).

IR(KBr)cm⁻¹ : 1705, 1695, 1520, 1400, 1345, 1110

NMR(CDCl₃) δ: 1.74(2H,m),2.0-2.60(4H,m),2.34(3H,s),3.0-3.5(3H,m), 3.84(1H,m),4.0-4.3(2H,m),5.22(2H,s),6.02(1H,br), 7.52(2H,d,J=9Hz) ,8.24(2H,d,J=9Hz) ##STR1333##

The same procedure as in Reference Example 5-6 was carried out by using the compound obtained by the above reaction (7 g, 17 mmol) to obtain the above identified compound (5.94 g, yield: 95.6%).

IR(KBr)cm⁻¹ : 1695, 1520, 1400, 1345, 1105

NMR(CDCl₃) δ: 1.5-1.88(2H,m),1.75(1H,d,J=8Hz),2.12(1H,dd,J=18,8Hz), 2.37(1H,dd,J=18,8Hz),2.44(1H,m),3.0-3.56(5H,m), 4.0-4.22(2H,m),5.22(2H,s),6.14(1H,br),7.52 (2H,d,J=9Hz),8.23(2H,d,J=9Hz)

REFERENCE EXAMPLE 20 (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[3-[(p-nitrobenzyloxycarbonyl)amino]-2-pyrrolidon-4-yl]pyrrolidine ##STR1334##

1.6M n-butyllithium (2.5 ml, 4 mmol) was dropwise added to a solution of diisopropylamine (0.64 ml, 4.6 mmol) in tetrahydrofuran (40 ml) under a nitrogen stream at -78° C. and the reaction solution was stirred for 45 minutes. A solution of (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonyl-2-pyrrolidon-4-yl)pyrrolidine (986 mg, 2 mmol, compound of Reference Example 5-1) in tetrahydrofuran (10 ml) was dropwise added to this solution, and the mixture was stirred at the same temperature for 2 hours. Then, p-toluenesulfonyl azide (945 mg, 4.8 mmol) was added thereto, and the mixture was stirred further for 1 hour. Then, trimethylsilyl chloride (1.29 ml, 10 mmol) was added to this reaction solution, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure. A saturated sodium hydrogen carbonate aqueous solution was added to the residue, and then the mixture was extracted with methylene chloride (50 ml). The organic layer was washed with a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, chloroform) to obtain (2S,4R)-2-(3-azido-N-tert-butoxycarbonyl-2-pyrrolidon-4-yl)-N-tert-butoxycarbonyl-4-(tert-butyldimethylsiloxy)pyrrolidine (720 mg, yield: 68.5%).

IR(KBr)cm⁻¹ : 2110, 1790, 1760, 1695, 1315, 1155

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.48(9H,s),1.6-2.2(2H,m), 3.1-3.85(5H,m),4.10(1H,br),4.26(1H,br),4.5(1H,br) ##STR1335##

The compound obtained by the above reaction (720 mg, 1.37 mmol) was dissolved in methanol (50 ml), and a 10% palladium-carbon catalyst (200 mg) was added thereto. The mixture was vigorously stirred under a hydrogen stream for 2 hours. The mixture was filtered, and the obtained filtrate was concentrated to obtain crude oil (730 mg). This oil was dissolved in a mixture of dioxane (20 ml) and water (20 ml). The pH was adjusted to 8 with triethylamine, and then 4,6-dimethyl-2-(p-nitrobenzyloxycarbonylthio)pyrimidine (504 mg, 1.6 mmol) was added thereto. This solution was stirred for 6 hours, while maintaining the pH at a level of 8. The reaction solution was concentrated under reduced pressure. The residue was extracted with ethyl acetate (50 ml). The extract was washed sequentially with a saturated sodium hydrogen carbonate aqueous solution and a saturated sodium chloride aqueous solution. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, chloroform-methanol 100:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[N-tert-butoxycarbonyl-3-[(p-nitrobenzyloxycarbonyl)amino]-2-pyrrolidon-4-yl]pyrrolidine (840 mg, yield: 90.6%).

IR(KBr)cm⁻¹ : 1795, 1695, 1525, 1350, 1255, 1155

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.48(9H,s),1.54(9H,s), 1.72-2.1(2H,m),3.08-3.78(5H,m),4.1-4.3(3H,m), 5.2(2H,br),7.5(2H,d,J=9Hz),8.24(2H,d,J=9Hz) ##STR1336##

The compound obtained by the above reaction (840 mg, 1.2 mmol) was dissolved in dry methylene chloride (20 ml), and the solution was cooled to 0° C. Then, trifluoroacetic acid (1 ml, 13 mmol) was added thereto, and the mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated, and trifluoroacetic acid was removed. Then, the residue was dissolved in methanol (30 ml), and a 2.5N hydrochloric acid-methanol solution (0.63 ml, 1.6 mmol) was added thereto, and the mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated. Then, the obtained residue was dissolved in a mixture of dioxane (20 ml) and water (20 ml). While adjusting the pH to a level of 8 with triethylamine, 4,6-dimethyl-2-(p-nitrobenzyloxycarbonylthio)pyrimidine (850 mg, 2.6 mmol) was added thereto, and the mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated, and the residue was extracted with ethyl acetate (100 ml). The organic layer was washed with a saturated sodium chloride aqueous solution. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, chloroform-methanol 50:1) to obtain (2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[3-[(p-nitrobenzyloxycarbonyl)amino]-2-pyrrolidon-4yl]pyrrolidine (450 mg, yield: 69%).

IR(KBr)cm⁻¹ : 1700, 1520, 1345

NMR(CDCl₃ +CD₃ OD) δ: 1.96-2.24(2H,m),3.06-3.84(5H,m),4.08-4.5(3H,m),5.22 (2H,s),5.24(2H,s),7.54(4H,d,J=9Hz),8.22(4H,d,J=9Hz) ##STR1337##

The same procedure as in Reference Example 5-4 was carried out by using the compound obtained by the above reaction (450 mg, 0.83 mmol) to obtain crude (2S,4R)-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)-2-[3-[(p-nitrobenzyloxycarbonyl)amino]-2-pyrrolidon-4yl]pyrrolidine (620 mg).

IR(KBr)cm⁻¹ : 1710, 1610, 1525, 1350, 1175 ##STR1338##

The same procedure as in Reference Example 5-5 was carried out by using the compound obtained by the above reaction (620 mg) to obtain (2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[3-[(p-nitrobenzyloxycarbonyl)amino]-2-pyrrolidon-4-yl]pyrrolidine (130 mg, yield: 26%).

IR(KBr)cm⁻¹ : 1710, 1700, 1610, 1525, 1350, 1110

NMR(CDCl₃) δ: 1.86-1.96(2H,m),2.34(3H,s),2.68(1H,br),3.0-3.4(4H,m), 4.84(1H,m),4.1-4.4(2H,m),5.2(4H,br),5.92(1H,br), 6.84(1H,br),7.54(4H,d,J=9Hz),8.22(4H,d,J=9Hz) ##STR1339##

The same procedure as in Reference Example 5-6 was carried out by using the compound obtained by the above reaction (130 mg, 0.21 mmol) to obtain the above identified compound i.e. (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[3-[(p-nitrobenzyloxycarbonyl)amino]-2-pyrrolidon-4-yl]pyrrolidine (110 mg, yield: 91%).

IR(KBr)cm⁻¹ : 1710, 1610, 1515, 1350

NMR(CDCl₃) δ: 1.6-1.9(2H,br),1.77(1H,d,J=8Hz),2.77(1H,br), 3.95-4.42(4H,m),4.05-4.2(3H,br),5.19(2H,s), 5.21(2H,s),5.9(1H,br),6.92(1H,br),7.54(4H,d,J=9Hz), 8.2(4H,d,J=9Hz)

REFERENCE EXAMPLE 21 (2S,4S),N-allylpxycarbonyl-2-iodometyl-4-tritylthiopyrrolidine ##STR1340##

A solution of methanesulfonyl chloride (20.5 ml, 265 mmol) in methylene chloride (20 ml) was dropwise added to a solution of (2S,4S)-N-allyloxycarbonyl-2-hydroxymethyl-4-tritylthiopyrrolidine (119 g, 259 mmol, compound of Reference Example 3-1 of Japanese Patent Application No. 192,093/1990) and triethylamine (40.3 ml, 289 mmol) in methylene chloride (1 l) under cooling with ice, and the mixture was stirred at the same temperature for 30 minutes. The reaction solution was washed sequentially with water (500 ml), a saturated sodium hydrogen carbonate aqueous solution (250 ml) and a saturated sodium chloride aqueous solution (250 ml), then dried over anhydrous sodium sulfate and concentrated. To the residue, ethyl acetate (30 ml) and diisopropyl ether (270 ml) were added, and the precipitated crystals were collected by filtration to obtain (2S,4S)-N-allyloxycarbonyl-2-methanesulfonyloxymethyl-4-tritylthiopyrrolidine (130.8 g, yield: 94%).

NMR(CDCl₃) δ: 1.8(1H,m),2.1(1H,m),1.7-2.4(3H,m),3.0(3H,s), 3.9(1H,m),4.1-4.6(4H,m),5.2-5.4(2R,m),5.9(1H,m), 7.2-7.6(15H,m) ##STR1341##

A solution comprising the compound obtained by the above reaction (130.8 g, 243 mmol), sodium iodide (180 g, 1.2 mol) and 2-butanone (1.3 l), was heated and stirred for 3 hours. The reaction solution was washed sequentially with water (300 ml), a 10% sodium thiosulfate aqueous solution (200 ml) and a saturated sodium chloride aqueous solution (500 ml), then dried over anhydrous sodium sulfate and concentrated. To the residue, diisopropyl ether (120 ml) and n-hexane (300 ml) were added, and the precipitated crystals were collected by filtration to obtain the above identified compound (123.7 g, yield: 89%).

NMR(CDCl₃) δ: 1.6(1H,m),2.1(1H,m),2.7-3.1(3H,m),3.3-3.6(3H,m), 4.5(2H,br s),5.25(2H,m),5.9(1H,m),7.2-7.6(15H,m)

REFERENCE EXAMPLE 22 (2R,4S)-N-allyloxycarbonyl-4-mercapto-2-(2-oxopyrrolidin-3-ylmethyl)pyrrolidine ##STR1342##

Lithium diisopropylamide (2.1M tetrahydrofuran solution, 6.45 ml, 13.5 mmol) was dropwise added to a solution of N-tert-butyldimethylsilyl-2-oxopyrrolidine (1.80 g, 9.03 mmol) in tetrahydrofuran (150 ml) at -78° C., and the mixture was stirred at the same temperature for 15 minutes. Then, a solution of (2S,4S)-N-allyloxycarbonyl-2-iodomethyl-4-tritylthiopyrrolidine (2.57 g, 4.5 mmol) in tetrahydrofuran (18 ml) was dropwise added thereto, and the mixture was stirred at the same temperature for 1 hour. The reaction solution was poured into a mixture of ethyl acetate (150 ml) and a 10% ammonium chloride aqueous solution (50 ml) for liquid separation. The organic layer was washed sequentially with a 10% sodium dihydrogen phosphate aqueous solution (100 ml) and a saturated sodium chloride aqueous solution (100 ml), then dried over anhydrous sodium sulfate and concentrated. The residue was dissolved in tetrahydrofuran (12 ml) and methanol (3 ml), and 6N hydrochloric acid (3 ml) was added thereto under cooling with ice. The mixture was stirred for 1 hour. Ethyl acetate (50 ml) was added to the reaction solution, and the organic layer was washed sequentially with a saturated sodium hydrogen carbonate aqueous solution (30 ml) and a saturated sodium chloride aqueous solution (30 ml), then dried over anhydrous sodium sulfate and concentrated. The residue was subjected to silica gel flash column chromatography (Wakogel™ C-300, 40 ml, ethyl acetate-n-hexane 3:1) to obtain (2R,4S)-N-allyloxycarbonyl-2-(2-oxopyrrolidin-3-ylmethyl)-4-tritylthiopyrrolidine (880 mg, yield: 37%).

NMR(CDCl₃) δ: 1.5(1H,m),1.7-2.3(6H,m),2.6-3.0(3H,m),3.34(2H,m), 3.66(1H,m),4.5(2H,br s),5.3(2H,m),5.9(2H,m), 7.2-7.6(15H,m) ##STR1343##

Trifluoroacetic acid (1.3 ml) and triethylsilane (0.42 ml, 2.63 mmol) were added to a solution of the compound obtained by the above reaction (1.32 g, 2.51 mmol) in methylene chloride (1.3 ml) under cooling with ice, and the mixture was stirred at room temperature for 30 minutes. The organic solvent was removed under reduced pressure, and ethyl acetate (50 ml) was added to the residue. The mixture was washed sequentially with a 1M phosphate buffer solution (pH 5.5, 30 ml×2) and a saturated sodium chloride aqueous solution (30 ml), then dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel flash column chromatography (Wakogel™ C-300, 40 ml, acetone-ethyl acetate 3:7) to obtain the above identified compound (650 mg, yield: 91%).

NMR(CDCl₃) δ: 1.5-1.7(2H,m),1.9-2.4(5H,m),2.65(1H,m),3.1-3.4 (4H,m),3.92(1H,m),4.1(1H,m),4.6(2H,br d,J=6Hz), 5.2-5.4(2H,m),5.95(2H,m)

REFERENCE EXAMPLE 23 (2R,4S)-N-allyloxycarbonyl-4-mercapto-2-(2-oxoazetidin-3-ylmethyl)pyrrolidine ##STR1344##

The same procedure as in Reference Example 22-1 was carried out by using N-tert-butyldimethylsilyl-2-oxoazetidine (650 mg, 3.5 mmol), lithium diisopropylamide (2.1M tetrahydrofuran solution, 3.34 ml, 7.0 mmol), and (2S,4S)-N-allyloxycarbonyl-2-iodomethyl-4-tritylthiopyrrolidine (1 g, 1.75 mmol) to obtain (2R,4S)-N-allyloxycarbonyl-2-(2-oxoazetidin-3-ylmethyl)-4-tritylthiopyrrolidine (220 mg, yield: 24%).

NMR(CDCl₃) δ: 1.5(1H,m),1.9-2.3(2H,m),2.7-3.1(4H,m),3.2(1H,m), 3.4(1H,t,J=5Hz),3.7(1H,m),4.5(2H,br s),5.3(2H,m), 5.62(1H,br s),5.9(1H,m),7.2-7.6(15H,m) ##STR1345##

The same procedure as in Reference Example 22-2 was carried out by using the compound obtained by the above reaction (510 mg, 0.99 mmol) and triethylsilane (0.17 ml, 1.05 mmol) to obtain the above identified compound, which was used for the subsequent reaction without purification.

REFERENCE EXAMPLE 24 (2R,4S)-N-allyloxycarbonyl-2-(N-allyloxycarbonyl-pyrrolidin-3-ylmethyl)-4-mercaptopyrrolidine ##STR1346##

Di-tert-butyl dicarbonate (1.97 ml, 8.6 mmol) was added to a solution comprising (2S,4S)-N-allyloxycarbonyl-2-[(Z)-2-oxopyrrolidin-3-ylidenemethyl]-4-tritylthiopyrrolidine (3 g, 5.7 mmol, compound of Reference Example 7-1 of Japanese Patent Application No. 192,093/1990), triethylamine (0.8 ml, 5.7 mmol), 4-(dimethylamino)pyridine (699 mg, 5.7 mmol) and tetrahydrofuran (30 ml). The mixture was stirred at room temperature for 5 hours, and then the solvent was removed under reduced pressure. Ethyl acetate (50 ml) was added to the residue, and the mixture was washed sequentially with a 10% sodium dihydrogen phosphate aqueous solution (20 ml) and a saturated sodium chloride aqueous solution (20 ml), then dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel flash column chromatography (Wakogel™ C-300, 40 ml, ethyl acetate-n-hexane 1:2) to obtain (2S,4S)-N-allyloxycarbonyl-2-[(Z)-N-tert-butoxycarbonyl-2-oxopyrrolidin-3-ylidenemethyl]-4-tritylthiopyrrolidine (3.24 g, yield: 91%).

NMR(CDCl₃) δ: 1.5(1H,m),1.56(9H,s),2.4-3.2(6H,m),3.7(2H,m), 4.3-4.6(2H,m),5.1-5.5(3H,m),5.7-6.0(2H,m), 7.2-7.6(15H,m) ##STR1347##

Lithium chloride (68 mg, 1.6 mmol), sodium borohydride (61 mg, 1.6 mmol) and ethanol (5 ml) were sequentially added to a solution comprising the compound obtained by the above reaction (500 mg, 0.8 mmol) and tetrahydrofuran (2.5 ml), and the mixture was stirred at room temperature for 16 hours. Ethyl acetate (30 ml) was added to the reaction solution, and the mixture was washed sequentially with water (15 ml×2) and a saturated sodium chloride aqueous solution (15 ml), then dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel flash column chromatography (Wakogel™ C-300, 40 ml, ethyl acetate-n-hexane 1:1) to obtain (2R,4S)-N-allyloxycarbonyl-2-[4-tert-butoxycarbonylamino-2-(hydroxymethyl)butyl]-4-tritylthiopyrrolidine (270 mg, yield: 54%).

NMR(CDCl₃) δ: 1.2-1.5(4H,m),1.44(9H,s),1.7-2.i(2H,m),2.22(1H,m), 2.6-3.0(3H,m),3.16(2H,m),3.4-3.8(3H,m),4.5(2H,br s), 4.7(0.5H,br s),4.9(0.5H,br s),5.2-5.4(2H,m), 5.9(1H,m),7.2-7.6(15H,m) ##STR1348##

Methanesulfonyl chloride (35 μl, 0.45 mmol) was dropwise added to a solution comprising the compound obtained by the above reaction (270 mg, 0.43 mmol), triethylamine (66 μl, 0.47 mmol) and methylene chloride (3 ml) under cooling with ice, and the mixture was stirred at the same temperature for 30 minutes. Methylene chloride (30 ml) was added to the reaction solution, and the mixture was washed sequantially with water (15 ml), a saturated sodium hydrogen carbonate aqueous solution (15 ml) and a saturated sodium chloride aqueous solution (15 ml), then dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel flash column chromatography (Wakogel™ C-300, 40 ml, ethyl acetate-n-hexane 1:1) to obtain (2R,4S)-N-allyloxycarbonyl-2-[4-tert-butoxycarbonylamino-2-(methanesulfonyloxymethyl)butyl]-4-tritylthiopyrrolidine (270 mg, yield: 89%).

NMR(CDCl₃) δ: 1.3-2.0(6H,m),1.44(9H,s),2.2(1H,m),2.6-3.3(5H,m), 3.0(1.5H,s),3.02(1.5H,s),3.8(1H,m),4.14(2H,m), 4.4-4.7(2.5H,m),5.02(0.5H,br s),5.2-5.4(2H,m), 5.9(1H,m),7.2-7.6(15H,m) ##STR1349##

The compound obtained by the above reaction (260 mg, 0.367 mmol) was dissolved in a 1.55N hydrogen chloride-methanol solution (0.7 ml), and the mixture was stirred at room temperature for 4 hours. The solvent was removed under reduced pressure, and then tetrahydrofuran (5 ml) and triethylamine (0.11 ml, 0.81 mmol) were added thereto, and the mixture was stirred at room temperature for 1 hour. Triethylamine (0.11 ml, 0.81 mmol) was added to the reaction solution, and then allyl chloroformate (43 μl, 0.4 mmol) was dropwise added thereto under cooling with ice. The mixture was stirred at the same temperature for 30 minutes. Ethyl acetate (30 ml) was added to the reaction solution, and the mixture was washed sequentially with water (15 ml), a saturated sodium hydrogen carbonate aqueous solution (15 ml) and a saturated sodium chloride aqueous solution (15 ml), then dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel flash column chromatography (Wakogel™ C-300, 40 ml, ethyl acetate-n-hexane 1:1) to obtain (2R,4S)-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-3-ylmethyl)-4-tritylthiopyrrolidine (180 mg, yield: 82%).

NMR(CDCl₃) δ: 1.3-1.5(3H,m),1.8-2.2(4H,m),2.6-3.0(4H,m),3.3(1H,m), 3.4-3.7(3H,m),4.5(2H,br s),4.6(2H,br d,J=5Hz),5.2-5.4 (4H,m),5.8-6.1(2H,m),7.2-7.6(15H,m) ##STR1350##

The same procedure as in Reference Example 22-2 was carried out by using the compound obtained by the above reaction (1.03 g, 1.73 mmol) and triethylsilane (0.29 ml, 1.81 mmol) to obtain the above identified compound (560 mg, yield: 92%).

NMR(CDCl₃) δ: 1.5-1.8(4H,m),1.9-2.3(4H,m),2.62(1H,m),2.9-4.2 (7H,m),4.6(4H,m),5.2-5.4(4H,m),5.9-6.1(2H,m)

REFERENCE EXAMPLE 25 (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-carbamoylpyrrolidin-4-ylmethyl]-4-mercaptopyrrolidine ##STR1351##

The same procedure as in Reference Example 22-1 was carried out by using N-tert-butyldimethylsilyl-5-tert-butyldimethylsiloxymethyl-2-oxopyrrolidine (10.67 g, 31.6 mmol), lithium diisopropylamide (2.1M tetrahydrofuran solution, 15.1 ml, 31.6 mmol) and (2S,4S)-N-allyloxycarbonyl-2-iodomethyl-4-tritylthiopyrrolidine (10 g, 17.6 mmol) to obtain (2R,4S)-N-allyloxycarbonyl-2-[(5S)-5-hydroxymethyl-2-oxopyrrolidin-3-ylmethyl]-4-tritylthiopyrrolidine (2 g, yield: 21%).

NMR(CDCl₃) δ: 1.3-2.4(8H,m),2.7-3.0(2H,m),3.5(1H,m),3.6-3.8(3H,m), 4.5(2H,br s),5.2-5.4(2H,m),5.9(1H,m),6.3(1H,br s), 7.2-7.6(15H,m) ##STR1352##

4-(dimethylamino)pyridine (88 mg, 0.72 mmol) and triethylamine (1.2 ml, 8.6 mmol) were sequentially added to a solution comprising the compound obtained by the above reaction (4 g, 7.18 mmol), tert-butyldimethylsilyl chloride (1.19 g, 7.9 mmol) and N,N-dimethylformamide (20 ml) under cooling with ice, and the mixture was stirred at room temperature for 3 hours. Ethyl acetate (80 ml) was added to the reaction solution, and the mixture was washed sequentially with water (40 ml×2), a saturated sodium hydrogen carbonate aqueous solution (40 ml) and a saturated sodium chloride aqueous solution (40 ml), then dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel flash column chromatography (Wakogel™ C-300, 40 ml, ethyl acetate-n-hexane 2:1) to obtain (2R,4S)-N-allyloxycarbonyl-2-[(5S)-5-tert-butyldimethylsiloxymethyl-2-oxopyrrolidin-3-ylmethyl]-4-tritylthiopyrrolidine (4.1 g, yield: 85%).

NMR(CDCl₃) δ: 0.05(6H,s),0.9(9H,s),1.3-2.4(8H,m),2.8-3.0(2H,m), 3.3-3.7(4H,m),4.5(2H,br s),5.2-5.4(2H,m),5.9(2H,m), 7.2-7.6(15H,m) ##STR1353##

The same procedure as in Reference Example 24-1 was carried out by using the compound obtained by the above reaction (4.1 g, 6.1 mmol), di-tert-butyl dicarbonate (2.53 ml, 11 mmol), 4-(dimethylamino)pyridine (746 mg, 6.1 mmol) and triethylamine (0.94 ml, 6.7 mmol) to obtain (2R,4S)-N-allyloxycarbonyl-2-[(5S)-N-tert-butoxycarbonyl-5-tert-butyldimethylsiloxymethyl-2-oxopyrrolidin-3-ylmethyl]-4-tritylthiopyrrolidine (4.7 g, yield: 100%).

NMR(CDCl₃) δ: 0.04(3H,s),0.06(3H,s),0.9(9H,s),1.5(2H,m),1.52 (9H,s),1.9(2H,m),2.15(2H,m),2.8-3.0(4H,m),3.62(1H,m), 3.7(1H,dd,J=10,2Hz),3.9(1H,dd,J=10,4Hz),4.1(1H,m), 4.5(2H,br s),5.1-5.3(2H,m),5.9(1H,m),7.2-7.6(15H,m) ##STR1354##

The same procedure as in Reference Example 24-2 was carried out by using the compound obtained by the above reaction (4.7 g, 6.1 mmol), lithium chloride (518 mg, 12.2 mmol) and sodium borohydride (463 mg, 12.2 mmol) to obtain (2R,4S)-N-allyloxycarbonyl-2-[(4S)-4-tert-butoxycarbonylamino-5-tert-butyldimethylsiloxy-2-hydroxymethylpentyl]-4-tritylthiopyrrolidine (2.18 g, yield: 46%).

NMR(CDCl₃) δ: 0.04(6H,s),0.9(9H,s),1.4-1.8(6H,m),1.48(9H,s), 2.2(1H,m),2.6-3.0(3H,m),3.5-3.9(6H,m),4.5(2H,br s), 4.7(1H,br s),5.2-5.4(2H,m),5.9(1H,m),7.2-7.6(15H,m) ##STR1355##

The same procedure as in Reference Example 24-3 was carried out by using the compound obtained by the above reaction (2.18 g, 2.8 mmol), methanesulfonyl chloride (0.24 ml, 3 mmol) and triethylamine (0.47 ml, 3.4 mmol) to obtain (2R,4S)-N-allyloxycarbonyl-2-[(4S)-4-tert-butoxycarbonylamino-5-tert-butyldimethylsiloxy-2-methanesulfonyloxymethylpentyl]-4-tritylthiopyrrolidine (2.15 g, yield: 90%).

NMR(CDCl₃) δ: 0.04(6H,s),0.9(9H,s),1.3-1.9(6H,m),1.45(9H,s), 2.2(1H,m),2.6-3.0(3H,m),3.0(3H,s),3.4-3.8(4H,m), 4.1-4.7(5H,m),5.1-5.3(2H,m),5.9(1H,m),7.2-7.6(15H,m) ##STR1356##

The same procedure as in Reference Example 24-4 was carried out by using the compound obtained by the above reaction (2.15 g, 2.52 mmol), a 1N hydrogen chloride-methanol solution (25 ml), allyl chloroformate (0.35 ml, 3.28 mmol) and triethylamine (1.76 ml, 12.6 ml) to obtain (2R,4S)-N-allyloxycarbonyl-2[(2S)-N-allyloxycarbonyl-2-hydroxymethylpyrrolidin-4-ylmethyl]-4-tritylthiopyrrolidine (1.21 g, yield: 77%).

NMR(CDCl₃) δ: 1.3-2.2 (7H,m),2.6-3.05(4H,m),3.5-3.7(4H,m),4.05 (1H,m),4.45(2H,br d,J=4Hz),4.6(2H,m),5.1-5.3(4H,m), 5.8-6.0(2H,m),7.2-7.6(15H,m) ##STR1357##

A solution comprising the compound obtained by the above reaction (1.21 g, 1.93 mmol), pyridinium dichromate (3.63 g, 9.65 mmol) and N,N-dimethylformamide (7.2 ml), was stirred at room temperature for 16 hours. Ethyl acetate (50 ml) was added to the reaction solution, and the mixture was washed with water (20 ml×2). To the organic layer, water (30 ml) containing potassium carbonate (270 mg, 1.93 mmol) was added for liquid separation. The aqueous layer was acidified with 6N hydrochloric acid, and ethyl acetate (50 ml) was added for liquid separation. The organic layer was washed with a saturated sodium chloride aqueous solution (20 ml), then dried over anhydrous sodium sulfate and concentrated to obtain (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-carboxypyrrolidin-4-ylmethyl]-4-tritylthiopyrrolidine (1 g, yield: 81%).

NMR(CDCl₃) δ: 1.3-1.5(2H,m),1.8-2.4(5H,m),2.7-3.1(4H,m),3.5-3.9 (2H,m),4.3-4.8(5H,m), 5.1-5.5(4H,m),5.8-6.0(2H,m), 7.2-7.6 (15H,m) ##STR1358##

Isobutyl chloroformate (90 μl, 0.69 mmol) was dropwise added to a solution comprising the compound obtained by the above reaction (400 mg, 0.62 mmol), triethylamine (96 μl, 0.69 mmol) and tetrahydrofuran (6 ml) at -15° C., and the mixture was stirred at the same temperature for 20 minutes. Then, concentrated aqueous ammonia (0.18 ml) was added thereto, and the mixture was stirred at 0° C. for 1 hour. Ethyl acetate (30 ml) was added to the reaction solution, and the mixture was washed sequentially with a saturated sodium hydrogen carbonate aqueous solution (15 ml) and a saturated sodium chloride aqueous solution (15 ml), then dried over anhydrous sodium sulfate and-concentrated to obtain (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-carbamoylpyrrolidin-4-ylmethyl]-4-tritylthiopyrrolidine (310 mg, yield: 78%).

NMR(CDCl₃) δ: 1.3-2.5(7H,m),2.7-3.1(4H,m),3.72(2H,m),4.3-4.7(5H,m), 5.1-5.5(5H,m),5.9-6.1(2.5H,m)6.8(0.5H,br s), 7.2-7.6(15H,m) ##STR1359##

The same procedure as in Reference Example 22-2 was carried out by using the compound obtained by the above reaction (310 mg, 0.48 mmol) and triethylsilane (81 μl, 0.51 mmol) to obtain the above identified compound (190 mg, yield: 100%).

NMR(CDCl₃) δ: 1.5-2.7(8H,m),3.0-3.3(3H,m),3.7(1H,m),3.86(1H,m), 4.06(1H,m),4.4(1H,d,J=8Hz),4.6(4H,m),5.2-5.5(5H,m), 5.8-6.0(2.5H,m),6.8(0.5H,br s)

REFERENCE EXAMPLE 26 (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-(methylcarbamoyl)pyrrolidin-4-ylmethyl]-4mercaptopyrrolidine ##STR1360##

The same procedure as in Reference Example 25-8 was carried out by using (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-carboxypyrrolidin-4-ylmethyl]-4-tritylthiopyrrolidine (300 mg, 0.47 mmol), isobutyl chloroformate (74 μl, 0.56 mmol), triethylamine (78 μl, 0.56 mmol) and a 40% methylamine aqueous solution (0.19 ml) to obtain (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-(methylcarbamoyl)pyrrolidin-4-ylmethyl]-4-tritylthiopyrrolidine (300 mg, yield: 98%).

NMR(CDCl₃) δ: 1.3-2.5(7H,m),2.6-3.0(3H,m),2.8(3H,d,J=5Hz), 3.6(2H,m),4.3(1H,d,J=8Hz),4.4-4.6(4H,m),5.1-5.4 (4H,m),5.8-6.0(2.5H,m),6.7(0.5H,br s),7.2-7.6(15H,m) ##STR1361##

The same procedure as in Reference Example 22-2 was carried out by using the compound obtained by the above reaction (300 mg, 0.46 mmol) and triethylsilane (83 μl, 0.5 mmol) to obtain the above identified compound (168 mg, yield: 89%).

NMR(CDCl₃) δ: 1.5-2.7(8H,m),2,82(3H,d,J=5Hz),3.0-3.3(3H,m),3.7 (1H,m),3.9(1H,m),4.1(1H,m),4.4(1H,d,J=8Hz),4.6(4H,m), 5.2-5.4(4H,m),5.8-6.1(2.5H,m),6.9(0.5H,br s)

REFERENCE EXAMPLE 27 (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-(dimethylcarbamoyl)pyrrolidin-4-ylmethyl]-4-mercaptopyrrolidine ##STR1362##

The same procedure as in Reference Example 25-8 was carried out by using (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl-2-carboxypyrrolidin-4-ylmethyl]-4-tritylthiopyrrolidine (300 mg, 0.47 mmol), isobutyl chloroformate (74 μl, 0.56 mmol), triethylamine (78 μl, 0.56 mmol) and a 50% dimethylamine aqueous solution (0.27 ml) to obtain (2R,4S)-N-allyloxycarbonyl-2-[(2S)-N-allyloxycarbonyl- 2-(dimethylcarbamoyl)pyrrolidin-4-ylmethyl]-4-tritylthiopyrrolidine (290 mg, yield: 93%).

NMR(CDCl₃) δ: 1.3-2.3(7H,m),2.6-3.0(4H,m),2.9(6H,s),3.5(2H,m), 4.1-4.7(5H,m),5.1-5.3(4H,m),5.7-5.9(2H,m), 7.2-7.6(15H,m) ##STR1363##

The same procedure as in Reference Example 22-2 was carried out by using the compound obtained by the above reaction (290 mg, 0.43 mmol) and triethylsilane (76 μl, 0.48 mmol) to obtain the above identified compound (110 mg, yield: 60%).

NMR(CDCl₃) δ: 1.5-2.7(8H,m),2.9-3.3(3H,m),2.98(3H,s),3.1(3H,s), 3.7-4.2(3H,m),4.4-4.8(5H,m),5.2-5.4(4H,m),5.9(2H,m)

REFERENCE EXAMPLE 28 (2S, 4S )-2-(2,4-dioxoimidazolidin-5-yl)-4-(p-methoxybenzylthio)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine ##STR1364##

Oxalyl chloride (1.3 ml, 14.9 mmol) was dropwise added to a solution of dimethyl sulfoxide (2.2 ml, 3.10 mmol) in methylene chloride (30 ml) under a nitrogen stream at -78° C. and the reaction solution was stirred at the same temperature for 30 minutes. A solution of (2S,4S)-2-hydroxymethyl-4-(p-methoxybenzylthio)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (4.32 g, 9.99 mmol) in methylene chloride (20 ml) cooled to -78° C., was dropwise added to this mixture. This mixture was stirred at the same temperature for 30 minutes, and then triethylamine (7.0 ml, 50.2 mmol) was dropwise added thereto. This mixture was stirred at the same temperature for 10 minutes and further at room temperature for 1 hour. Then, methylene chloride (400 ml) was added thereto, and the mixture was washed with a 1N potassium hydrogen sulfate aqueous solution and water. The organic layer was dried over anhydrous sodium sulfate. Then, solvent was distilled off to obtain a crude aldehyde.

The crude aldehyde obtained by the above reaction was dissolved in a solvent mixture comprising ethanol (15 ml), water (15 ml) and N,N-dimethylformamide (6 ml). Then, ammonium carbonate (4.50 g, 46.8 mmol) and then sodium cyanide (0.98 g, 20.0 mmol) were added thereto, and the mixture was stirred at 60° C. for 6 hours. The mixture was cooled to room temperature. Then, the organic solvent was distilled off, and water (100 ml) and chloroform (100 ml) were added to the residue. The organic layer was separated, and then the aqueous layer was extracted with chloroform (100 ml×2). The organic layers were put together, then washed sequentially with water and a saturated sodium chloride aqueous solution and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography (Wakogel™ C-300, methanol-chloroform) to obtain the above identified compound (1.82 g, yield: 36%).

IR(KBr)cm⁻¹ : 3250, 1770, 1730, 1710, 1610, 1510, 1350

NMR(CDCl₃ +CD₃ OD) δ: 1.9-2.3(2H,m),3.7-4.1(1H,m),4.26(1H,m),4.72 and 5.04 (1H,s),5.22(2H,s),6.86(2H,d,J=8Hz),7.24(2H,d,J=8Hz), 7.51(2H,d,J=8Hz),8.26(2H,d,J=8Hz)

REFERENCE EXAMPLE 29 (2R,4S)-N-allyloxycarbonyl-2-(2,4-dioxoimidazolidin-5-ylmethyl)-4-tritylthiopyrrolidine diastereomers A and B ##STR1365##

A 1.6M n-butyllithium-hexane solution (4.1 ml, 6.56 mmol) was dropwise added to a solution of diisopropylamine (1 ml, 7.14 mmol) in tetrahydrofuran (5 ml) under a nitrogen stream at -78° C., and the reaction solution was stirred at the same temperature for 10 minutes and under cooling with ice for 30 minutes. A solution of N,N'-bis(tert-butyldimethylsilyl)hydantoin (1.65 g, 5.02 mmol) in tetrahydrofuran (2 ml) was dropwise added to this reaction solution at -78° C. This solution was stirred at the same temperature for 1 hour. Then, hexamethylphosphoric triamide (1.75 ml, 10 mmol) was added to this reaction solution, and the mixture was stirred for 10 minutes Then, a solution of (2S,4S)-N-allyloxycarbonyl-2-iodomethyl-4-tritylthiopyrrolidine (2.28 g, 4.00 mmol, compound of Reference Example 21-2) in tetrahydrofuran (3 ml) was dropwise added thereto. The reaction solution was stirred at the same temperature for 2 hours. To the reaction solution, a saturated ammonium chloride aqueous solution (10 ml) and ethyl acetate (300 ml) were added. The organic layer was washed sequentially with a 1N potassium hydrogen sulfate aqueous solution, water and a saturated sodium chloride aqueous solution and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate) to obtain (2R,4S)-N-allyloxycarbonyl-2-(1-tert-butyldimethylsilyl-2,4-dioxoimidazolidin-5-ylmethyl)-4-tritylthiopyrrolidine (1.19 g, yield: 45%).

IR(KBr)cm⁻¹ : 3420, 3250, 1765, 1700, 1400, 1200

NMR(CDCl₃) δ: 0.27 and 0.35(6H,s),0.94 and 0.97(9H,s),1.5(1H,m), 1.7(1H,m),2.25(1H,m),2.6-3.0(3H,m),4.2(1H,m), 4.3-4.6(3H,m),5.25(2H,m},6.85(1H,m),7.1-7.7(15H,m), 8.06 and 8.11(1H,s) ##STR1366##

A mixture of a 49% hydrofluoric acid (1.5 ml) and acetonitrile (13.5 ml) was added to a solution of the compound obtained by the above reaction (1.10 g, 1.68 mmol) in acetonitrile (15 ml). This solution was stirred overnight at room temperature. Ethyl acetate (300 ml) was added to this reaction solution. This solution was washed sequentially with water, a saturated sodium hydrogen carbonate aqueous solution, water and a saturated sodium chloride aqueous solution and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography (Wakogel™ C-300, methanol-chloroform) to obtain diastereomer A (550 mg, yield: 61%, polar compound) and diastereomer B (220 mg, yield: 24%, low polar compound) of the above identified compound.

Diastereomer A

IR(KBr)cm⁻¹ : 3550, 3480, 3420, 1770, 1730, 1680, 1445, 1410

NMR(CDCl₃) δ: 1.2-2.4(4H,m),2.75(2H,m),3.05(1H,m),3.8(1H,m),4.10 (1H,d,J=8Hz),4.49(2H,d,J=6Hz),5.26(2H,m),5.9(1H,m), 6.54(1H,s),7.0-7.7(15H,m),8.41(1H,s)

Diastereomer B

IR(KBr)cm⁻¹ : 3420, 3230, 1775, 1730, 1700, 1445, 1410

NMR(CDCl₃) δ: 1.2-1.9(2H,m),1.95-2.45(2H,m),2.7(2H,m),3.05(1H,m), 3.8-4.05(2H,m),4.46(2H,d,J=5Hz),5.26(2H,m), 5.85(1H,m),6.59(1H,s),7.1-7.6(15H,m),8.12(1H,s)

REFERENCE EXAMPLE 30 (2R,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-(2,5-dioxopyrrolidin-3-ylmethyl)pyrrolidine ##STR1367##

Ethyl 3-[(2S,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonylpyrrolidin-2-yl]acrylate (10 g, 26.8 mmol) was subjected to catalytic hydrogenation in ethanol (200 ml) by means of 10% palladium carbon (500 mg) at 60° C. for 3 hours. The reaction solution was left to cool, and the catalyst was filtered off. The filtrate was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (Wakogel™ C-300, 300 ml, ethyl acetate-hexane 1:10) to obtain ethyl 3-[(2R,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonylpyrrolidin-2-yl]propionate (9.91 g, yield: 98%).

NMR(CDCl₃) δ: 0.04(6H,s),0.90(9H,s),1.28(3H,t,J=8Hz),1.48(9H,s), 1.78(2H,m),2.00(2H,m),2.30(2H,m),3.38(2H,m), 3.96(1H,m),4.14(2H,q,J=8Hz),4.32(1H,m) ##STR1368##

2.1M lithium diisopropylamide (7.6 ml, 16.0 mmol) was dropwise added to a solution of the compound obtained by the above reaction (5.0 g, 13.3 mmol) in tetrahydrofuran (250 ml) in a nitrogen stream at -78° C., and the mixture was stirred for 10 minutes. Then, allyl bromide (3.46 ml, 40 mmol) was dropwise added thereto, and the mixture was stirred at the same temperature for 30 minutes. After removing the cooling medium, the mixture was stirred for 30 minutes. Then, a saturated ammonium chloride aqueous solution was added thereto, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and then concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate-hexane 1:6) to obtain 2-ethoxycarbonyl-1-[(2R,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonylpyrrolidin-2yl]-4-pentene (6.09 g, yield: 110%).

NMR(CDCl₃) δ: 0.08(6H,s),0.86(9H,s),1.26(3H,t,J=8Hz),1.48(9H,s), 1.50-2.20(3H,m),2.16-2 74(2H,m),3.34(2H,m), 3.90 (1H,m),4.14(2H,m),4.32 (1H,m),5.04(2H,m), 5.70(1H,m) ##STR1369##

Ruthenium trichloride (6.25 mg, 30 μmol) was added to a mixture comprising the compound obtained by the above reaction (507 mg, 1.22 mmol), carbon tetrachloride (2.5 ml), acetonitrile (2.5 ml), sodium periodate (1.1 g, 5.14 mmol) and water (3.75 ml). The reaction solution was stirred for 2 hours and then extracted with methylene chloride. The organic layer was dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate-hexane 1:1) to obtain 3-ethoxycarbonyl-4-[(2R,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonylpyrrolidin-2-yl]butyric acid (480 mg, yield: 90%).

NMR(CDCl₃) δ: 0.08(6H,s),0.88(9H,s),1.28(3H,t,J=8Hz),1.48(9H,s), 1.78(2H,m),2.04(2H,m),2.80(3H,m),3.38(2H,m), 3.90(1H,m),4.20(2H,m),4.36(1H,m) ##STR1370##

Carbonyldiimidazole (475 mg, 2.93 mmol) was added to a solution of the compound obtained by the above reaction (847 mg, 1.95 mmol) in tetrahydrofuran (17 ml) at room temperature, and the reaction solution was stirred for 1 hour. Then, concentrated aqueous ammonia (3.4 ml) was added thereto. The reaction solution was stirred further for 30 minutes and then diluted with ethyl acetate. The organic layer was washed with a saturated ammonium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated. The residue was dissolved in tetrahydrofuran (15 ml). Then, 60% sodium hydride (94 mg, 2.3 mmol) was added thereto under cooling with ice, and the mixture was stirred for 20 minutes. Then, the mixture was subjected to liquid separation treatment as described above, and the organic layer was dried and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate-hexane 1:1) to obtain (2R,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonyl-2-(2,5-dioxopyrrolidin-3-ylmethyl)pyrrolidine (576 mg, yield: 76%).

NMR(CDCl₃) δ: 0.08(6H,s),0.90(9H,s),1.48(6H,s),1.70(2H,m), 2.00(2H,m),2.58(1H,m),2.90(3H,m),3.38(2H,m), 4.12(1H,m),4.40(1H,m) ##STR1371##

The compound obtained by the above reaction (570 mg, 1.47 mmol) was dissolved in a mixture of methanol (11 ml) and thionyl chloride (0.32 ml, 4.4 mmol) under cooling with ice. The reaction solution was returned to room temperature and stirred for 30 minutes, and it was then concentrated under reduced pressure. The residue was dissolved in chloroform (10 ml). Then, triethylamine (0.61 ml, 5.86 mmol) and 4,6-dimethyl-2-(p-nitrobenzyloxycarbonylthio)pyrimidine (470 mg, 1.47 mmol) were added thereto, and the mixture was stirred overnight. The reaction solution was washed with 1N hydrochloric acid, then dried over anhydrous magnesium sulfate and concentrated. The residue was dissolved in methylene chloride (10 ml). Then, methanesulfonyl chloride (0.14 ml, 1.77 mmol) and triethylamine (0.31 ml, 2.2 mmol) were added thereto, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was poured into a saturated sodium hydrogen carbonate aqueous solution and extracted with methylene chloride. The organic layer was dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate) to obtain (2R,4R)-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)-2-(2,5-dioxopyrrolidin-3-ylmethyl)pyrrolidine (420 mg, yield: 62%).

NMR(CDCl₃) δ: 1.80-2.40(4H,m),2.45-3.30(5H,m),3.06(3H,s), 4.00-4.40(2H,m),5.26(2H,m),7.54(2H,d,J=8Hz), 8.28(2H,d,J=8Hz) ##STR1372##

A mixture comprising the compound obtained by the above reaction (410 mg, 0.90 mmol), N,N-dimethylformamide (4.1 ml) and potassium thioacetate (308 mg, 2.70 mmol) was stirred at 70° C. for 1 hour. The reaction solution was poured into water and extracted with ethyl acetate. The organic layer was dried and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate-hexane 1:1) to obtain (2R,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-(2,5-dioxopyrrolidin-3-ylmethyl)pyrrolidine (251 mg, yield:

NMR(CDCl₃) δ: 1.70(2H,m),2.16(2H,m),2.38(3H,s),2.40-3.40(5H,m), 3.80-4.30(2H,m),5.22(2H,m),7.56(2H,d,J=8Hz), 8.24(2H,d,J=8Hz)

REFERENCE EXAMPLE 31 (2S,4S)-4-acetylthio-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)pyrrolidine ##STR1373##

Triethylamine (1.27 ml, 9.55 mmol) was added to a solution of (2S,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonylprolinal (19.1 g, 57.87 mmol) in nitromethane (95 ml) under stirring and cooling with ice. This solution was left to stand at room temperature for hours. The solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography (Wakogel™ C-300, 350 g, hexane-ethyl acetate 9:1) to obtain oily (2S,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonyl-2-(1-hydroxy-2-nitroethyl)pyrrolidine (14.81 g, yield: 65.5%). IR(KBr)cm⁻¹ : 3420, 1675, 1700, 1560, 1415, 1255, 1165, 840, 780

NMR(CDCl₃) δ: 0.07(6H,s),0.87(9H,s),1.49(9H,s),1.7-2.1(2H,m), 3.2-3.6(2H,m),4.12-4.56(5H,m) ##STR1374##

Thionyl chloride (3.58 ml, 49.33 mmol) was dropwise added to a solution of the compound obtained by the above reaction (14.8 g, 37.89 mmol) in methylene chloride (150 ml) at -50° C. The reaction solution was stirred at -50° C. for 5 minutes, and then triethylamine (16.5 ml, 118.39 mmol) was dropwise added thereto. The cooling bath was removed, and the reaction solution was stirred at room temperature for 3 hours, then poured into ice water (100 ml) and extracted with methylene chloride. The organic layer was washed with water and a 10% sodium hydrogen carbonate aqueous solution, then dried over anhydrous sodium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 95:5) to obtain (2S,4R)-4-tert-butyldimethylsiloxy-N-tert-butoxycarbonyl-2-(2-nitrovinyl)pyrrolidine (11.48 g, yield: 81.3%) as solid.

IR(KBr)cm⁻¹ : 1685, 1520, 1400, 1350, 1250, 1160, 835, 770

NMR(CDCl₃) δ: 0.09(6H,s),0.80(9H,s),1.46(9H,br s),1.80-2.28(2H,m), 3.40-3.71(2H,m),4.36-4.76(2H,m),7.02-7.24(2H,m) ##STR1375##

Triethylamine (0.74 ml, 5.58 mmol) was added to a mixture comprising the compound obtained by the above reaction (2.0 g, 5.37 mmol), methylene chloride (40 ml), tetrahydrofuran (10 ml) and glycine ethyl ester hydrochloride (750 mg, 5.37 mmol) at room temperature, and the mixture was stirred for 30 minutes, then poured into a saturated sodium hydrogen carbonate aqueous solution and extracted with methylene chloride. The organic layer was dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate-hexane 1:3) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[1-(ethoxycarbonylmethyl)amino-2-nitroethyl]pyrrolidine (2.32 g, yield: 91%).

NMR(CDCl₃ ) δ: 0.08(6H,s),0.88(9H,s),1.28(3H,t,J=6Hz),1.50 (9H,br s),1.98(2H,m),3.24(1H,m),3.50(2H,m), 3.80(1H,m),4.08(1H,m),4.18(1H,m),4.20(2H,q,J=6Hz), 4.40(3H,m) ##STR1376##

A mixture of the compound obtained by the above reaction (1.26 g, 2.65 mmol), ammonium formate (1.67 g, 26.6 mmol) and methanol (25 ml) was treated with 10% palladium carbon (300 mg) at room temperature for 1 hour, then at 60° C. for 1 hour. The reaction mixture was filtrated to remove palladium carbon and concentrated. The residue was dissolved in methylene chloride and washed with water. The organic layer was dried over anhydrous magnesium sulfate and concentrated to obtain an oily substance containing (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(5-oxopiperazin-2-yl)pyrrolidine.

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.48(9H,br s),1.96(2H,m), 3.22(3H,m),3.52(3H,m),4.10(1H,m),4.32(1H,m), 4.38(1H,m) ##STR1377##

Allyl chlorocarbonate (0.33ml, 3.18 mmol) and triethylamine (0.55 ml, 3.98 mmol) were added to a solution of the compound obtained by the above reaction in methylene chloride (25 ml) under cooling with ice, and the mixture was stirred for 1 hour. The reaction solution was poured into a saturated sodium hydrogen carbonate aqueous solution, then extracted with methylene chloride, dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate-hexane 3:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)pyrrolidine diastereomer A (low polar compound, 710 mg, yield (2 steps): 59%) and diastereomer B (highly polar compound, 290 mg, yield (2 steps): 24%).

Diastereomer A

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.46(9H,br s),1.86(2H,m), 3.20-3.90(5H,m),4.10(1H,m),4.38(1H,m),4.48(2H,m), 4.64(2H,m),5.30(2H,m),5.90(1H,m)

Diastereomer B

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.44(9H,br s),1.82(1H,m), 2.00(1H,m),3.10-3.80(4H,m),4.00-4.50(5H,m), 4.60(2H,m),5.30(2H,m),5.90(1H,m) ##STR1378##

Methylene chloride (4.5 ml) and trifluoroacetic acid (4.5 ml) were added to the compound obtained by the above reaction (diastereomer A, 993 mg, 2.05 mmol) under cooling with ice, and the mixture was returned to room temperature, then stirred for 10 minutes and concentrated under reduced pressure. Then, methylene chloride (18 ml) and triethylamine (1.51 ml, 10.3 mmol) were added thereto, and a solution of allyl chlorocarbonate (0.69 ml, 6.16 mmol) in methylene chloride (2.8 ml) was dropwise added thereto under cooling with ice. The mixture was stirred for 1 hour, then poured into a saturated sodium hydrogen carbonate aqueous solution and extracted with methylene chloride. The organic layer was dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate) to obtain (2S,4R)-N-allyloxycarbonyl-4-tert-butyldimethylsiloxy-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)pyrrolidine diastereomer A (698 mg, yield: 73%).

The same operation was carried out by using the diastereomer B obtained by the above reaction (726 mg, 1.50 mmol) to obtain (2S,4R)-N-allyloxycarbonyl-4-tert-butyldimethylsiloxy-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)pyrrolidine diastereomer B (451 mg, yield: 64%).

Diastereomer A

NMR(CDCl₃) δ: 0.04(6H,s),0.86(9H,s),1.86(2H,m),3.50(4H,m), 3.80(1H,m),4.16(1H,m),4.44(3H,m),4.58(2H,m), 4.66(2H,m),5.32(4H,m),5.96(2H,m)

Diastereomer B

NMR(CDCl₃) δ: 0.04(6H,s),0.86(9H,s),1.86(1H,m),2.02(1H,m), 3.20-3.80(4H,m),4.20(3H,m),4.44(2H,m),4.60(4H,m), 5.30(4H,m),5.92(2H,m) ##STR1379##

A 1M tetrabutylammonium fluoride solution (2.28 ml, 2.28 mmol) was added to a solution of the compound obtained by the above reaction (diastereomer A, 698 mg, 1.49 mmol) in tetrahydrofuran (7.0 ml) under cooling with ice, and the mixture was stirred at room temperature for 2 hours. The reaction solution was diluted with ethyl acetate, then washed with a saturated sodium hydrogen carbonate aqueous solution, dried over anhydrous magnesium sulfate and concentrated. The residue was dissolved in methylene chloride (14 ml), and triethylamine (0.88 ml, 5.98 mmol) and methanesulfonyl chloride (0.24 ml, 2.99 mmol) were added thereto under cooling with ice. The mixture was stirred for 40 minutes. The reaction solution was poured into a sodium hydrogen carbonate aqueous solution, then extracted with methylene chloride, dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate) to obtain (2S,4R)-N-allyloxycarbonyl-4-methanesulfonyloxy-2-(1-allyloxycarbonyl-5-oxopiperazin- 2-yl)pyrrolidine diastereomer A (487 mg, yield: 75%).

The same operation was carried out by using the diastereomer B obtained by the above reaction (451 mg, 0.96 mmol) to obtain (2S,4R)-N-allyloxycarbonyl-4-methanesulfonyloxy-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)pyrrolidine diastereomer B (285 mg, yield: 68%).

Diastereomer A

NMR(CDCl₃) δ: 2.24(2H,m),3.08(3H,s),3.56(2H,m),3.58(1H,m), 3.72(2H,m),4.18(2H,m),4.50(2H,m),4.64(4H,m), 5.36(4H,m),5.86(2H,m)

Diastereomer B

NMR(CDCl₃) δ: 2.18(1H,m),2,48(1H,m),3.04(3H,s),3.20-3.80(4H,m), 3.90-4.30(4H,m),4.62(5H,m),5.30(4H,m),5.92(2H,m) ##STR1380##

A mixture comprising the compound obtained by the above reaction (diastereomer A, 487 mg, 1.12 mmol), potassium thioacetate (386 mg, 3.38 mmol) and N,N-dimethylformamide (9.7 ml), was stirred at 70° C. for 1.5 hours. The mixture was left to cool, then diluted with ethyl acetate, washed with a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate) to obtain (2S,4S)-N-allyloxycarbonyl-4-acetylthio-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)pyrrolidine diastereomer A (508 mg, yield: 109%).

The operation was conducted in the same manner as above by using the diastereomer B obtained by the above reaction, to obtain (2S,4S)-N-allyloxycarbonyl-4-acetylthio-2-(1-allyloxycarbonyl-5-oxopiperazin-2-yl)pyrrolidine diastereomer B (200 mg, yield: 73%).

Diastereomer A

NMR(CDCl₃) δ: 1.78(1H,m),2.36(3H,s),2.44(1H,m),3.10(1H,m), 3.52(2H,m),3.82(2H,m),4.36(4H,m),4.66(4H,m), 5.34(4H,m),5.98(2H,m)

Diastereomer B

NMR(CDCl₃) δ: 1.80(1H,m),2.64(1H,m),2.38(3H,s),3.30(1H,m), 3.68(2H,m),3.96(1H,m),4.10-4.60(5H,m),4.62(4H,m), 5.28(4H,m),5.98(2H,m)

REFERENCE EXAMPLE 32 (2S,4S)-4-mercapto-N-(p-nitrobenzyloxycarbonyl)-2-[N-p-nitrobenzyloxycarbonyl)piperidin-4-yl]pyrrolidine ##STR1381##

To a mixture of cuprous iodide (50 mg, 0.26 mmol) and a 1M vinylmagnesium bromide tetrahydrofuran solution (15 ml) in tetrahydrofuran (100 ml), a mixture of trimethylsilyl chloride (3.18 ml, 25.0 mmol) and (E)-3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]acrylic acid ethyl ester (2.0 g, 5.0 mmol) in tetrahydrofuran (30 ml) was dropwise added over 30 minutes in a nitrogen stream at -78° C. The reaction mixture was stirred for 2 hours at the same temperature, then quenched with a saturated ammonium chloride aqueous solution (15 ml) and extracted with ethyl acetate (100 ml). The organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution, then dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 15:1) to obtain 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-5-pentenoic acid ethyl ester (1.7 g, yield: 79.4%).

NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,S),1.24(3H,t,J=8Hz),1.48(9H,s), 1.66-1.97(2H,m),2.12-2.59(2H,m),3.10-3.30(2H,m), 3.33-3.70(1H,m),3.93-4.24(3H,m),4.30(1H,m), 5.08(1H,d,J=10Hz),5.11(1H,d,J=18Hz),5.64(1H,m) ##STR1382##

To a solution of lithium aluminum hydride (113 mg, 2.98 mmol) in tetrahydrofuran (30 ml), the compound obtained by the above reaction (1.7 g, 3.98 mmol) in tetrahydrofuran (10 ml) was added in a nitrogen stream at 0° C., and the reaction mixture stirred for 1 hour at the same temperature. The reaction mixture was quenched with a saturated ammonium chloride aqueous solution (5 ml) and extracted with ethyl acetate (100 ml). The organic layer was successively washed with a 1N sodium hydroxide aqueous solution, water and a saturated sodium chloride aqueous solution, dried over sodium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 5:1) to obtain 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin- 2-yl]pent-4-en-1-ol (1.09g, yield: 59.9%).

NMR(CDCl₃) δ: 0.04(6H,s),0.86(3H,s),1.46(9H,s),1.50-1.95(4H,m), 2.73(1H,m),3.12(1H,dd,J=12,4Hz),3.32-3.77(3H,m), 3.88-4.20(1H,m),4.32(1H,m),5.12(2H,m),5.64(1H,m) ##STR1383##

Triphenylphosphine (1.0 g, 3.81 mmol) and phthalimide (0.63 g, 4.28 mmol) were added to a stirred solution of the compound obtained by the above reaction (1.09 g, 2.83 mmol) in tetrahydrofuran (30 ml) under a nitrogen stream, and then diethyl azodicarboxylate (0.58 ml, 3.68 mmol) added thereto under cooling with ice. The reaction mixture was stirred for 2 hours at the same temperature, then extracted with ethyl acetate (70 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C.-300, hexane-ethyl acetate 6:1) to obtain 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl ]-5-phthalimido-1-pentene (1.31 g, yield: 90.0%).

NMR(CDCl₃) δ: 0.03(6H,s),0.84(9H,s),1.42(9H,s),1.50-1.93(4H,m), 2.73(1H,m),3.20(1H,dd,J=12,4Hz),3.28-3.80(3H,m), 3.84-4.17(1H,m),4.28(1H,m),5.04-5.30(2H,m), 5.62(1H,m),7.71(2H,m),7.84(2H,m) ##STR1384##

To a solution of the compound obtained by the above reaction (1.31 g, 2.55 mmol) in ethanol (15 ml) was added hydrazine monohydrate (0.37 ml, 7.63 mmol) at room temperature. The reaction mixture was stirred overnight at the same temperature, then concentrated under reduced pressure. The residue was taken up in water and ethyl acetate. The organic layer was successively washed with a 2N ammonium hydroxide aqueous solution, water and a saturated sodium chloride aqueous solution, then dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was dissolved in tetrahydrofuran (30 ml) and then di-tert-butyl dicarbonate (640 mg, 2.93 mmol) added thereto at room temperature. After being stirred for 1 hour, the reaction mixture was concentrated under reduced pressure and the residue subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 5:1) to obtain 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-5-tert-butoxycarbonylamino-1-pentene (983 mg, yield: 79.3%).

NMR(CDCl₃) δ: 0.04(6H,s),0.86(9H,s),1.42(9H,s),1.46(9H,s), 1.82(2H,m),2.48-2.82(1H,m),3.00(1H,m),3.22(2H,m), 3.37-3.73(1H,m),4.01(1H,m),4.30(1H,m),4.42-4.75 (1H,m),5.02-5.22(2H,m),5.58(1H,m) ##STR1385##

To a solution of 9-borabicyclo[3.3.1]nonane (545 mg, 2.23 mmol) in tetrahydrofurane (10 ml) was added a solution of the compound obtained by the above reaction (980 mg, 2.02 mmol) in tetrahydrofuran (5 ml) in a nitrogen stream at room temperature and the reaction mixture stirred for 2.5 hours at the same temperature. Water (6 ml) and sodium perborate tetrahydrate (1.23 g, 7.99 mmol) were added thereto. The resulting mixture was stirred vigorously overnight at room temperature, and then extracted with ethyl acetate (50 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 2:1) to obtain 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-5-tert-butoxycarbonylamino-1-propanol (753 mg, yield: 71.5%).

NMR(CDCl₃) δ: 0.06(6H,s),0.87(9H,s),1.44(9H,s),1.48(9H,s), 2.30(1H,m),3.00-3.40(4H,m),3.54(1H,m),3.74(2H,m), 4.08(1H,m),4.30(1H,m) ##STR1386##

The same procedure as in Reference Example 13-4 was carried out by using the compound obtained by the above reaction (753 mg, 1.50 mmol), potassium tert-butoxide (370 mg, 3.30 mmol) and p-toluenesulfonyl chloride (315 mg, 1.65 mmol) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonylpiperidin-4-yl)pyrrolidine (494 mg, yield: 68.0%).

NMR(CDCl₃) δ: 0.05(6H,s),0.87(9H,s),1.00-1.28(2H,m),1.45(18H,s), 1.78(2H,m),2.64(2H,m),3.22(1H,dd,J=12,4Hz),3.38-3.66 (1H,m),3.96(1H,m),4.14(2H,m),4.29(1H,m) ##STR1387##

The same procedure as in Reference Example 5-3 was carried out by using the compound obtained by the above reaction (494 mg, 1.02 mmol) and 4,6-dimethyl-2-(p-nitrobenzyloxycarbonylthio)pyrimidine (684 mg, 2.14 mmol) to obtain (2S,4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidin-4-yl]pyrrolidine (485 mg, yield: 89.8%).

NMR(CDCl₃) δ: 1.05-1.34(2H,m),1.42-1.70(2H,m),1.94(2H,m), 2.13-2.46(2H,m),2.58-2.94(2H,m),3.38(1H,m),3.76 (1H,m),4.03-4.36(3H,m),4.45(1H,m),5.12-5.37(4H,m), 7.54(4H,d,J=8Hz),8.23(2H,d,J=8Hz),8.24(2H,d,J=8Hz) ##STR1388##

The same procedure as in Reference Example 5-4 was carried out by using the compound obtained by the above reaction (485 mg, 0.92 mmol) and methanesulfonyl chloride (82 μl, 1.06 mmol) to obtain (2S,4R)-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidin-4-yl]pyrrolidine (556 mg, yield: 99.9%).

NMR(CDCl₃) δ: 1.04-1.32(2H,m),1.44-1.75(2H,m),1.93-2.38(3H,m), 2.78(2H,m),3.05(3H,s),3.48(1H,m),4.04-4.35(4H,m), 5.12-5.38(5H,m),7.53(2H,d,J=8Hz),7.55(2H,d,J=8Hz), 8.25(2H,d,J=8Hz),8.26(2H,d,J=8Hz) ##STR1389##

The same procedure as in Reference Example 13-9 was carried out by using the compound obtained by the above reaction (556 mg, 0.92 mmol), potassium thioacetate (210 mg, 1.84 mmol) and sodium iodide (155 mg, 1.03 mmol) to obtain (2S,4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)piperidin-4-yl]pyrrolidine (476 mg, yield: 88.5%).

NMR(CDCl₃) δ: 1.07-1.35(2H,m) ,1.45-1.84(3H,m),2.10-2.46(5H,m), 2.76(2H,m), 3.00(1H,t,J=10Hz), 3.80(1H,m),3.98(1H,m), 4.26(3H,m), 5.23(4H,s),7.52(2H,d,J=8Hz),7.54 (2H,d,J=8Hz),8.23(2H,d,J=8Hz),8.24(2H,d,J=8Hz) ##STR1390##

The same procedure as in Reference Example 1-9 was carried out by using the compound obtained by the above reaction to obtain the above identified compound (433 mg, yield: 98%).

REFERENCE EXAMPLE 33 (2S,4S)-2-[2-carbamoyl-N-(p-nitrobenzyoxycarbonyl)pyrrolidin-4-yl]-4-mercapto-N-(p-nitrobenzyloxycarbonyl)pyrrolidine diastereomer III ##STR1391##

The same procedure as in Reference Example 6-2 was carried out by using (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-pyrrolidon-4-yl)pyrrolidine diastereomer B (10.0 g, 26.0 mmol, compound of Reference Example 12) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonyl-2-pyrrolidon-4-yl)pyrrolidine diastereomer B (12.37 g, yield: 98.1%).

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.46(9H,s),1.52(9H,s), 1.66(1H,m),1.96(1H,m),2.27(1H,dd,J=18,10Hz), 2.50(1H,dd,J=18,8Hz),3.24(1H,dd,J=13,4Hz), 3.62(1H,dd,J=13,8Hz),3.82(1H,dd,J=10,8Hz), 4.09(1H,m),4.32(1H,m) ##STR1392##

The same procedure as in Reference Example 13-2 was carried out by using the compound obtained by the above reaction (12.3 g, 25.4 mmol) to obtain 5-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-6-tert-butoxycarbonylamino-1-hexen-3-one diastereomer B (10.39 g, yield: 79.8%).

NMR(CDCl₃) δ: 0.04(6H,s),0.85(9H,s),1.42(9H,s),1.47(9H,s), 1.68(1H,m),1.92(1H,m),2.24-2.57(2H,m),3.16 (1H,dd,J=12,4Hz),3.23-3.68(2H,m),4.03(1H,m), 4.33(1H,m),5.82(1H,br d,J=10Hz),6.20(1H,br d,J=18Hz), 6.38(1H,dd,J=18,10Hz) ##STR1393##

The same procedure as in Reference Example 13-3 was carried out by using the compound obtained by the above reaction (10.3 g, 20.1 mmol) to obtain 5-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-6-tert-butoxycarbonylamino-1-hexen-3-ol (2.84 g, yield: 27.5%).

NMR(CDCl₃) δ: 0.04(6H,s),0.86(9H,s),1.44(9H,s),1.46(9H,s),1.73 (1H,m),1.95(1H,m),2.95(1H,m),3.18(1H,dd,J=12,4Hz), 3.56(2H,m),3.94(1H,m),4.30(1H,m),5.10(1H,d,J=10Hz), 5.30(1H,d,J=18Hz),5.86(1H,m) ##STR1394##

The same procedure as in Reference Example 13-4 was carried out by using the compound obtained by the above reaction (2.84 g, 5.52 mmol) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonyl-2-vinylpyrrolidin-4-yl)pyrrolidine (1.04 g, yield: 37.9%).

NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,s),1.42(9H,s),1.46(9H,s), 1.66-2.34(4H,m),3.02-3.34(2H,m),3.30-3.84(2H,m), 3.86-4.28(2H,m),4.34(1H,m),5.08(2H,m),5.76(1H,m) ##STR1395##

The same procedure as in. Reference Example 13-5 was carried out by using the compound obtained by the above reaction (1.04 g, 2.1 mmol ) to obtain (2S,4R)-N-tert-butoxycarbonyl- 4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonyl-2-carboxypyrrolidin-4-yl)pyrrolidine (500 mg, yield: 46.4%).

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.46(18H,s),1.68-2.50(5H,m), 3.26(2H,m),3.38-3.80(2H,m),3.90-4.40(3H,m) ##STR1396##

The same procedure as in Reference Example 9-3 was carried out by using the compound obtained by the above reaction (500 mg, 0.97 mmol) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-tert-butoxycarbonyl-2-carbamoylpyrrolidin-4-yl)pyrrolidine (388 mg, yield: 77.7%).

NMR(CDCl₃) δ: 0.02(6H,s),0.84(9H,s),1.42(18H,s),1.64-2.42(5H,m), 3.25(2H,m),3.36-3.80(2H,m),3.88-4.35(3H,m) ##STR1397##

The same procedure as in Reference Example 5-3 was carried out by using the compound obtained by the above reaction (388 mg, 0.76 mmol) to obtain (2S,4R)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (360 mg, yield: 85.4%).

NMR(CDCl₃) δ: 1.72-2.50(5H,m),3.22-3.55(2H,m),3.62-3.92(2H,m), 4.04-4.35(2H,m),4.46(1H,m),5.24(4H,m),7.52(4H,m), 8.20(4H,m) ##STR1398##

The same procedure as in Reference Example 5-4 was carried out by using the compound obtained by the above reaction (360 mg, 0.65 mmol) to obtain (2S,4R)-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (229 mg, yield: 55.8%).

NMR(CDCl₃) δ: 1.72-2.64(5H,m),3.07(3H,s),3.26-3.65(2H,m), 3.84(1H,m),4.08-4.36(3H,m),5.26(5H,m),7.52(4H,m), 8.18(4H,m). ##STR1399##

The same procedure as in Reference Example 13-9 was carried out by using the compound obtained by the above reaction (229 mg, 0.36 mmol) to obtain (2S,4S)-4-acetylthio-2-[2-carbamoyl-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-4-yl]-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (191 mg, yield: 86.5%).

NMR(CDCl₃ -CD₃ OD) δ: 1.58-1.88(2H,m),2.28(3H,s),2.48(3H,m),3.10(1H,m), 3.38(1H,m),3.78(2H,m),4.00(1H,m),4.09(2H,m),5.15 (4H,br s),7.46(4H,m),8.10(4H,br d,J=8Hz) ##STR1400##

The same procedure as in Reference Example 1-9 was carried out by using the compound obtained by the above reaction (191 mg, 0.31 mmol) to obtain the above identified compound (175 mg, 98.3%).

REFERENCE EXAMPLE 34 (2S,4S)-4-acetylthio-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-2-yl)pyrrolidine diastereomer A ##STR1401##

To a solution of propargyl 2-tetrahydropyranyl ether (15.2 g, 109 mmol) in tetrahydrofuran (165 ml), 1.6 N n-butyllithium in hexane (68.1 ml, 109 mmol) was added in a nitrogen stream under cooling with ice, and the mixture was stirred under cooling with ice for 1 hour. The mixture was cooled to -78° C. and added to a solution of (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyl-dimethylsiloxy-2-formylpyrrolidine (31.7 g, 90.2 mmol in tetrahydrofuran (150 ml) obtained by Reference Example 1-3) in a nitrogen stream at -78° C. and the mixture was stirred for 1 hour at the same temperature. After the reaction mixture was raised to -20° C. and stirred at the same temperature for 1.5 hours, a saturated ammonium chloride aqueous solution (150 ml) was added thereto and stirred. Ethyl acetate was added hereto and the organic layer was washed with water a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 1:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[1-hydroxy-4-(2-tetrahydropyranyloxy)-2-butynyl]pyrrolidine (38.2 g, yield: 90%).

IR(KBr)cm⁻¹ : 3400,2950,1700,1680,1400

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.46(9H,s),1.1-2.2(8H,m), 3.34(1H,m),3.43-3.63(2H,m),3.83(1H,m),4.04-4.54(5H, m),4.80(1H,br s) ##STR1402##

To a solution of the compound obtained by the above reaction (17.5 g, 37.2 mmol) in methanol (300 ml), 10% palladium-carbon catalyst (7 g) was added, and mixture was stirred in a hydrogen stream at room temperature for 3 hours. 10% palladium-carbon catalyst (7 g) was further added thereto and the mixture was stirred in a hydrogen stream at room temperature for 4 hours. The catalyst was filtered off and the filtrate was concentrated under reduced pressure to obtain (2S, 4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[1-hydroxy-4-(2-tetrahydropyranyloxy)butyl]pyrrolidine (14.8 g, yield: 84%). ##STR1403##

To a solution of the compound obtained by the above reaction (19.0 g, 40.1 mmol) in ethanol (420 ml), pyridinium p-toluenesulfonate (1.27 g, 4.75 mmol) was added, and the mixture was stirred at 55° C. for 1.5 hours. The reaction mixture was concentrated under reduced pressure and ethyl acetate was added thereto, washed sequentially with a saturated sodium hydrogen-carbonate aqueous solution, water and a saturated sodium chloride aqueous solution and dried over anhydrous magnesium sulfate. The solvent was concentrated under reduced pressure to obtain (2S,4R)-N-tern-butoxy-carbonyl-4-tert-butyldimethylsiloxy-2-[1,4-dihydroxybutyl)pyrrolidine (15.4 g, yield: 99%).

IR(KBr)cm⁻¹ : 3400,2950,1670,1400

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.46(9H,s),1.3-2.1(6H,m), 2.64(2H,br s),3.26(1H,m),3.4-3.8(4H,m),3.8-4.2(1H, m),4.32(1H,m) ##STR1404##

The compound obtained by the above reaction (2.42 g, 6.21 mmol) and p-toluenesulfonyl chloride (1,20 g, 6.29 mmol) were dissolved in pyridine, and the solution was stirred at room temperature for 1 hour. Diethyl ether was added to the reaction solution, and the precipitate was filtered off. The filtrate was washed sequentially with water, 10% citric acid, a saturated sodium hydrogencarbonate aqueous solution, water and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, heptane-ethyl acetate 1:i) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(1-hydroxy-4-p-toluenesulfonyloxybutyl)pyrrolidine (2.08 g, yield: 62%). ##STR1405##

To a solution of the compound obtained by the above reaction (2.08 g, 3.82 mmol) in dimethylsulfoxide (63 ml), sodium azide (0.90 g, 14 mmol) was added, and the mixture was stirred at 70° C. for 3 hours. Ethyl acetate was added thereto, washed with water, dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain (2S,4R)-2-(4-azido-1-hydroxy-butyl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidine (1.42 g, yield: 90%).

IR(KBr)cm⁻¹ : 3450,2100,1700,1680,1410

NMR(CDCl₃) δ; δ: 0.06(6H,s),0.86(9H,s),1.46(9H,s),1.5-2.1(7H,m),3.2-4.4(7H,m) ##STR1406##

To a solution of dimethyl sulfoxide (0,724 ml, 10.5 mmol) in methylene chloride (13 ml), oxalyl chloride (0.472 ml, 5.41 mmol) was dropwise added in a nitrogen stream at -78° C., and the reaction solution was stirred at the same temperature for 45 minutes. To the mixture was dropwise added a solution of the azide compound (1.21 g, 2.92 mmol) in methylene-chloride (4 ml) at -78° C. After the mixture was stirred at the same temperature for 30 minutes, triethylamine (2.44 ml, 17.5 mmol) was added thereto and the mixture was stirred at room temperature for 20 minutes. The mixture was washed sequentially with water, dilute hydrochloric acid, a saturated sodium hydrogencarbonate aqueous solution and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, heptane-ethyl acetate 2:1) to obtain (2S,4R)-2-(4-azidobutylyl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidine (1.02 g, yield: 85% ).

IR(KBr)cm⁻¹ : 2100,1700,1400

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.46(9H,s),1.7-2.2(4H,m), 2.62(2H,m),3.34(2H,m),3.56(2H,m),4.3-4.6(2H,m) ##STR1407##

To a solution of the compound obtained by the above reaction (2.34 g, 5.67 mmol) in methanol (60 ml), 10% palladium-carbon catalyst (236 mg) was added, and the mixture was stirred in a hydrogen stream an room temperature for 1 hour. 10% Palladium-carbon catalyst 236 mg) was added and hydrogenation was carried out at room temperature for 1 hour followed by further adding 10% palladium-carbon catalyst and hydrogenating for 2 hours. The catalyst was filtered off and to the filtrate were added acetic acid (0.67 ml, 11.7 mmol) and sodium cyanoborohydride (736 mg, 11.7 mmol) under cooling with ice, and the mixture was stirred at the same temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in chloroform, washed with a saturated sodium hydrogencarbonate aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, chloroform-methanol 97:3) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(pyrrolidin-2-yl)pyrrolidine (883 mg, yield: 42%).

NMR(CDCl₃) δ: 0.08(6H,s),0.86(9H,s),1.47 and 1.48(total 9H, both s),1.5-1.8(2H,m),1.9-2.4(4H,m),3.1-4.2(6H, m),4.32(1H,br s) ##STR1408##

To a solution of the compound obtained by the above reaction (883 mg, 2.38 mmol) in pyridine (4.5 ml), a solution of allyl chlorocarbonate (0.47 ml, 4.43 mmol) in methylene chloride (2.2 ml) was added under cooling with ice, and the mixture was stirred overnight at room temperature. Ethyl acetate was added to the reaction solution, washed sequentially with water, 10% citric acid, a saturated sodium hydrogencarbonate aqueous solution and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfane and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, heptane-ethyl acetate 2:1) to obtain (2S,4R)-2-(N-allyloxycarbonylpyrrolidin-2-yl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidine (718 mg, yield: 66%). ##STR1409##

To a solution of the compound obtained by the above reaction (912 mg, 2.00 mmol) and 2,6-lutidine (0.47 ml, 4.1 mmol) in methylene chloride (16 ml), trimethylsilyltrifluoromethanesulfonate (0.60 ml, 3.0 mmol) was dropwise added, and the mixture was stirred at the same temperature for 1.5 hours. After methanol (2.4 ml) was added to the reaction mixture and stirred at room temperature for 45 minutes, the solvent was distilled off under reduced pressure. The residue was dissolved in methylene chloride (16 ml) and to the solution was added triethylamine (0.61 ml, 4.38 mmol) followed by allyl chlorocarbonate (0.426 ml, 4.02 mmol) under cooling with ice, and the mixture was stirred at the same temperature for 1 hour, then at room temperature for 30 minutes. The reaction solution was washed sequentially with dilute hydrochloric acid, a saturated sodium hydrogencarbonate aqueous solution, water and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 2:1) to obtain (2S,4R)-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-2-yl)-4-tert-butyldimethylsiloxypyrrolidine diastereomer A (less polar compound; 330 mg, yield: 38%) and diastereomer B (polar compound; 242 mg, yield: 28%).

Diastereomer A

NMR(CDCl₃) δ: 0.05(6H,s),0.85(9H,s),1.88(6H,m),3.2-3.8(5H,m), 4.13(1H,m),4.42(1H,m),4.59(4H,m),5.1-5.4(4H,m),5.8-6.1(2H,m)

Diastereomer B

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.90(6H,m),3.2-3.7(5H,m),4.0-4.7(2H,m),4.58(4H,m),5.1-5.5(4H,m),5.8-6.1(2H,m) ##STR1410##

To a solution of diastereomer A obtained by the above reaction (330 mg, 0.752 mmol) in tetrahydrofuran (4 ml), 1 M tri-n-butylammonium fluoride in tetrahydrofuran (0.83 ml, 0.83 mmol) was added under cooling with ice, and the mixture was stirred at the same temperature for 45 minutes. Ethyl acetate was added to the reaction solution, washed sequentially with a saturated sodium hydrogencarbonate aqueous solution, water, a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was dissolved in methylene chloride (7 ml) and to the solution was added triethylamine (0.44 ml, 3.0 mmol) followed by methanesulfonyl chloride (0.12 ml, 1.5 mmol) under cooling with ice, and the mixture was stirred at the same temperature for 1 hour. The reaction solution was washed with a saturated sodium hydrogencarbonate agueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography Wakogel™ C-300, hexane-ethyl acetate 2:1) to obtain 2S,4R)-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-2-yl)-4-methanesulfonyloxypyrrolidine diastereomer A (249 mg, yield: 82%).

NMR(CDCl₃) δ: 1.89(4H,m),2.24(2H,m),3.04(3H,s),3.36(1H,m),3.54 (2H,m),3.9-4.4(3H,m),4.62(4H,m),5.2-5.5(5H,m),5.8-6.1(2H,m) ##STR1411##

To a solution of diastereomer A obtained by the above reaction (249 mg, 0.619 mmol) in N,N-dimethylformamide (2.7 ml), potassium thioacetate (141 mg, 1.24 mmol) and sodium iodide (92.9 mg, 0.619 mmol) were added, and the mixture was stirred at 70° C. for 4 hours. After cooling, ethyl acetate was added to the reaction solution, washed with water and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 2:1) to obtain the above identified compound (186 mg, yield: 79%).

NMR(CDCl₃) δ: 1.84(5H,m),2.34(3H,s),2.46(1H,m),3.01(1H,t,J=10Hz), 3.36(1H,m),3.58(1H,m),3.79(1H,m),4.23(3H,m),4.61 (4H,d,J=6Hz),5.1-5.5(4H,m),5.8-6.1(2H,m)

REFERENCE EXAMPLE 35 (2S,4S)-4-acetylthio-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-2-yl)pyrrolidine diastereomer B ##STR1412##

The same procedure as in Reference Example 34-10) was carried out by using (2S,4R)-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidin-2-yl)-4-tert-butyldimethylsiloxypyrrolidine diastereomer B (242 mg, 0.552 mmol, the compound of Reference Example 34-9) to obtain (2S, 4R)-N-allyloxycarbonyl-2-(N-allyloxycarbonylpyrrolidine 2-yl)-4-methanesulfonyloxypyrrolidine diastereomer B (172 mg, yield: 78%).

NMR(CDCl₃) δ: 1.5-2.5(6H,m),3.04(3H,S),3.2-4.6(6H,m),4.60(4H, br s),5.1-5.5(5H,m),5.8-6.1(2H,m) ##STR1413##

The same procedure as in Reference Example 34-11) was carried out by using the compound obtained by the above reaction (172 mg, 0.428 mmol) to obtain the above identified compound (133 mg, yield: 81%).

NMR(CDCl₃) δ: 1.64(2H,m),1.93(3H,m),2.34(3H,s),2.47(1H,m),3.0-3.8(3H,m),3.86(1H,m),4.0-4.5(3H,m),4.60(4H,br 5.1-5.5(4H,m),5.8-6.1(2H,m)

REFERENCE EXAMPLE 36 (2S,4S)-4-acetylthio-N-allyloxycarbonyl-2-(1-allyloxycarbonyl-5-oxopiperazin-3-yl)pyrrolidine ##STR1414##

To a mixture of (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-vinylpyrrolidine (4.0 g, 13.2 mmol), acetone (40 ml), water (40 ml) and N-methylmorpholin-N-oxide (2.5 g, 20 mmol), 4% osmium tetraoxide aqueous solution (2.0 ml) was added, and the mixture was stirred overnight at room temperature. After removing acetone under reduced pressure, the reaction solution was adjusted to pH 2.0 with 6 N sulfuric acid and extracted with ethyl acetate. The organic layer was dried and concentrated to obtain oily substance (4.35 g, yield: 98%) containing (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-1,2-dihydroxyethyl)pyrrolidine.

NMR(CDCl₃) δ: 0.06(6H,s),0.82(9H,s),1.40(9H,s),2.00(2H,m),3.28 (2H,m),3.52(2H,m),3.98(1H,m),4.30(2H,m) ##STR1415##

To a solution of the compound obtained by the above reaction (8.7 g, 26.0 mmol) in methylene chloride (174 ml), methanesulfonyl chloride (6.0 ml, 78 mmol) and triethylamine (16.2 ml, 116 mmol) were added under cooling with ice, and the mixture was stirred for 1 hour. The reaction solution was washed with a saturated sodium hydrogencarbonate aqueous solution, dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 1:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[1,2-di(methanesulfonyloxy)ethyl]pyrrolidine (9.50 g, yield: 74.5%).

NMR(CDCl₃) δ: 0.06(6H,s),0.84(9H,s),1.46(9H,br s),2.00(2H,m), 3.04(3H,s),3.08(3H,s),3.36(2H,m),4.10(1H,m),4.30 (1H,m),4.36(2H,m),5.40(1H,m) ##STR1416##

A mixture of the compound obtained by the above reaction (9.50 g, 19.3 mmol), sodium azide (8.72 g, 134 mmol) and dimethylsulfoxide (95 ml) was stirred at 100° C. for 1.5 hours. After cooling, ethyl acetate was added to the reaction mixture, washed sequentially with water and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 5:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(1,2-diazidoethyl)pyrrolidine (4.54 g, yield: 61%).

NMR(CDCl₃) δ: 0.06(6H,s),0.88(9H,s),1.48(9H,br s),2.00(2H,m), 3.10-3.90(5H,m),4.26(1H,m),4.40(1H,m) ##STR1417##

A mixture of the compound obtained by the above reaction (720 mg, 1.87 mmol), 10% palladium-carbon catalyst (140 mg) and methanol (14 ml) was stirred in a hydrogen stream for 2 hours. The catalyst was filtered off and the filtrate was concentrated. The residue was dissolved in methylene chloride (14 ml) and to the solution was added 4,6-dimethyl-2-(benzyloxycarbonylthio)pyrimidine under cooling with ice, then the mixture was stirred for 30 minutes. Acetoxyacetyl chloride (0.32 ml, 2.99 mmol) and triethylamine (0.52 ml, 3.74 mmol) were added thereto, and the mixture was stirred for 1 hour. The reaction solution was washed with a saturated sodium hydrogencarbonate aqueous solution, dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 2:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[1-(α-acetoxyacetamido)-2(benzyloxycarbonylamino)ethyl]pyrrolidine (102 g, yield: 9.6%). NMR(CDCl₃) δ: 0.06(6H,s),0.88(9H,s),1.86(1H,m),2.06(1H,m),2.20 (3H,s),3.10-3.70(4H,m),3.80(1H,m),4.16(1H,m),4.34 (1H,m),4.46 and 4.58(2H,ABq,J=16Hz),5.14(2H,br s), 7.38(5H,m) ##STR1418##

A mixture of the compound obtained by the above reaction (1.41 g, 2.49 mmol), 10% palladium-carbon catalyst (280 mg) and methanol (28 ml) was stirred in a hydrogen stream at room temperature for 6 hours. The catalyst was filtered off and the filtrate was concentrated under reduced pressure. The residue was treated with p-toluenesulfonyl chloride in pyridine at room temperature. After 4 hours, pyridine was distilled off under reduced pressure and the residue was dissolved in ethyl acetate. The solution was washed with a saturated sodium hydrogencarbonate aqueous solution, dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 1:3) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[1-(α-acetoxyacetamido)-2-(p-toluenesulfonylamino)ethyl]pyrrolidine (984 mg, yield: 67%).

NMR(CDCl₃) δ: 0.08(6H,s),0.88(9H,s),1.46(9H,s),1.76(1H,m),2.10 (1H,m),2.26(3H,s),2.44(3H,s),2.80-4.60(9H,m),5.70 (1H,br t,J=4Hz),7.32(2H,d,J=8Hz),7.76(2H,d,J=8H) ##STR1419##

A solution of the compound obtained by the above reaction (1.16 g, 1.97 mmol) in methanol (22 ml. was treated with 2 N sodium hydroxide aqueous solution (1.48 ml, 2.96 mmol) under cooling with ice for 15 minutes. The reaction solution was poured into a sodium chloride aqueous solution, extracted with methylene chloride, dried over anhydrous magnesium sulfate and concentrated. A solution of the obtained residue in methylene chloride (20 ml) was treated with triethylamine (0.33 ml, 2.36 mmol) and methanesulfonyl chloride (0.18 ml, 2.36 mmol) under cooling with ice for 30 minutes. The reaction solution was poured into a sodium hydrogen-carbonate aqueous solution, extracted with methylene chloride, dried over anhydrous magnesium sulfate and concentrated. To a solution of the obtained residue in methylene chloride (20 ml), DBU (1,8-diazabicyclo[5.4.0 ]undec-7-ene) (0.35 ml, 2.36 mmol) was added under cooling with ice, and the mixture was stirred at room temperature for 1 hour. The reaction solution was washed with a sodium hydrogencarbonate aqueous solution, dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 1:2) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[1-(p-toluenesulfonyl)-5-oxo-piperazin-3-yl]pyrrolidine (855 mg, yield: 82%).

NMR(CDCl₃) δ: 0.08(6H,s),0.86(9H,s),1.46(9H,s),1.92(1H,m),2.48 (3H,s),2.72-4.40(9H,m),7.40(2H,d,J=8Hz),7.70(2H,d, J=8Hz) ##STR1420##

The compound obtained by the above reaction (850 mg, 1.61 mmol) was treated with sodium metal in liquid ammonia at -78° C. Sodium metal was added until blue of the reaction solution didn't disappear, and the solution was stirred for 1 hour. To the reaction mixture was added solid ammonium chloride to neutralize excess sodium. The mixture was raised to room temperature in a nitrogen stream and ammonia was removed. To the residue was added water, extracted with ethyl acetate, dried over anhydrous magnesium sulfate and concentrated. The obtained residue was dissolved in methylene chloride (15 ml), and the solution was treated with triethylamine (0.34 ml, 2.41 mmol) and allyl chloroformate (0.20 ml, 1.93 mmol) under cooling with ice for 1 hour. The reaction solution was washed with a sodium hydrogencarbonate aqueous solution, dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromaography (Wakogel™

C-300, hexane-ethyl acetate 3:1) to obtain less polar compound (133 mg, yield: 18%) and polar compound (410 mg, yield: 55%) of (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(1-allyloxycarbonyl-5-oxopiperazin-3-yl)pyrrolidine.

Less polar compound

NMR(CDCl₃) δ: 0.08(6H,s),0.86(9H,s),1.48(9H,s),1.80(2H,m),3.10-4.50(9H,m),4.64(2H,m),5.20-5.40(2H,m),5.80-6.04(1H,m)

Polar compound

NMR(CDCl₃) δ: 0.08(6H,S),0.86(9H,s,)2.00(2H,m),3.00-4.50(9H,m), 4.64(2H,m),5.20-5.40(2H,m),5.80-6.04(1H,m) ##STR1421##

To the polar compound obtained by the above reaction (410 mg, 0.90 mmol), methylene chloride (4.0 ml) and trifluoroacetic acid (4.0 ml) were added, and the mixture was stirred at room temperature for 30 minutes. After concentrating under reduced pressure, the residue was dissolved in methylene chloride (8.0 ml) and triethylamine (0.38 ml, 2.73 mmol) and allyl chlorocarbonate (0.11 ml, 1.1 mmol) were added thereto, then the mixture was stirred for 1 hour. The reaction solution was washed with a saturated sodium hydrogen-carbonate aqueous solution, dried over anhydrous magnesium sulfate and concentrated to obtain oily substance (417 mg, yield: 105%) containing (2S,4R]-N-allyloxycarbonyl-4-tert-butyldimethylsiloxy-2-(1-allyloxycarbonyl-5-oxopiperazin-3-yl)pyrrolidine.

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.80(1H,m),2.04(1H,m),3.10-4.40(9H,m),4.64(4H,m),5.20-5.40(2H,m),5.84-6.02(2H,m) ##STR1422##

To a solution of the compound obtained by the above reaction (417 mg, 0.94 mmol) in tetrahydrofuran ml), 1 N tetrabutylammonium fluoride (0.94 ml, 0.94 mmol) was added, and the mixture was stirred at room temperature for 3 hours. The reaction solution was diluted with ethyl acetate, washed with a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated. The residue was dissolved methylene chloride (8.0 ml) and to the solution were added methanesulfonyl chloride (0.15 ml, 1.89 mmol) and triethylamine (0.39 ml, 2.83 mmol) under cooling with ice, then the mixture was stirred for 1 hour. The reaction solution was washed with a saturated sodium hydrogencarbonate aqueous solution, dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 5% methanol-ethyl acetate) to obtain (2S,4R)-N-allyloxycarbonyl-4-methanesulfonyloxy-2-(1-allyloxycarbonyl-5-oxopiperazin-3-yl)pyrrolidine (234 mg, yield: 61%).

NMR(CDCl₃) δ: 1.78(1H,m),2.08(1H,m),2.52(1H,m),3.06(3H,s),3.20-4.20(7H,m),4.66(4H,m),5.20-5.34(5H,m),5.80-6.04(2H,m) ##STR1423##

A mixture of the compound obtained by the above reaction (234 mg, 0.58 mmol), potassium thioacetic acid (197 ml, 1.73 mmol) and N,N-dimethylformamide (5.0 ml) was stirred at 70° C. for 1 hour. After cooling, the reaction solution was diluted with ethyl acetate, washed with a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, ethyl acetate) to obtain the above identified compound (168 mg, yield: 75%).

NMR(CDCl₃) δ: 1.78(1H,m),2.28(3H,s),2.58(1H,m),3.28(1H,m),3.50-4.40(8H,m),4.64(4H,m),5.22-5.40(4H,m),5.92(2H,m)

REFERENCE EXAMPLE 37 (2S,4S)-4-mercapto-2-(N-methylpiperidin-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine trifluoroacetic acid salt ##STR1424##

To a solution of 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-5-phthalimido-1-pentene (25 g, 48.5 mmol, compound of Reference Example 32-3) in methylene chloride (75 ml), trifluoroacetic acid (75 ml) was added under ice-cooling with stirring. After the mixture was stirred at the same temperature for 2 hours, the solvent and trifluoroacetic acid were distilled off under reduced pressure. Triethylamine (90 ml) was added to a solution of the residue in tetrahydrofuran (100 ml) under ice-cooling with stirring and a solution of n-nitrobenzyloxycarbonyl chloride (10.5 g, 48.5 mmol) in tetrahydrofuran (20ml) was dropwise added thereto. After the mixture was stirred at the same temperature for 1 hour, the solvent was distilled off under reduced pressure and ethyl acetate (200 ml) was added thereto. The organic layer was washed sequentially with water, 1N hydrochloric acid, a 5% sodium hydrogencarbonate aqueous solution and a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, heptane-ethyl acetate 2:1) to obtain 3-[(2S,4R)-4-tert-butyldimethylsiloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-2-yl]-5-phthalimido-1-pentene (23.47 g, yield: 81.5%).

NMR(CDCl₃) δ: 0.04(6H,s),0.82(9H,s),1.50-2.00(4H,m),2.70(1H,m), 3.38(1H,m),3.50-3.78(3H,m),4.12(1H,m),4.32(1H,m), 5.0-5.30(4H,m),5.60(1H,m),7.50(2H,br d,J=8Hz),7.68-7.90(4H,m),8.24(2H,br d,J=8Hz) ##STR1425##

To a solution of the compound obtained by the above reaction (23.47 g, 39.5 mmol) in ethanol (250 ml), hydrazine hydrate (5.82 ml, 0.12 mol) was added, the mixture was stirred at room temperature for 16 hours. The solvent was distilled off under reduced pressure, then ethyl acetate (200 ml) was added thereto. The organic layer was washed sequentially with 8% aqueous ammonia and a saturated sodium chloride agueous solution, then dried over anhydrous sodium sulfate and concentrated. Di-tert-butyl dicarbonate (8.72 q, 40 mmol) was added to a solution of the residue tetrahydrofuran (200 ml) at room temperature, and the mixture was stirred for 2 hours. The solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography (Wakogel™ C-300, heptane-ethyl acetate 2:1) to obtain 5-tert-butoxycarbonylamino-3-[(2S,4R)-tert-butyldimethylsiloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-2-yl]-1-pentene (18.9 g, yield: 85.0%).

NMR(CDCl₃) δ: 0.03(3H,s),0.05(3H,s),0.83(9H,s),1.43(9H,s),1.3-1.6 (2H,m),1.7-1.95(2H,m),2.8-3.4(3H,m),3.6(1H,m),4.13 (1H,m),4.36(1H,m),4.98-4.35(4H,m),4.58(1H,m),7.55 (2H,d,J=8Hz),8.25(2H,d,J=8Hz) ##STR1426##

To a solution of 9-borabicyclo[3.3.1]nonane (6.14 g, 50.3 mmol) in tetrahydrofuran (100 ml), a solution of the compound obtained by the above reaction (18.9 g, 33.5 mmol) in tetrahydrofuran (100 ml) was added in a nitrogen stream at room temperature, and the mixture was stirred for 3.5 hours. To the reaction solution, water (150 ml) and sodium perborate tetrahydrate (31.0 g, 0.2 mol) were added and the mixture was stirred an room temperature for 2 hours. Then the reaction solution was extracted with ethyl acetate (200 ml). The organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and then concentrated. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, heptane-ethyl acetate 2:1) to obtain 5-tert-butoxycarbonylamino-3-[(2S,4R)-4-tert-butyldimethyl-siloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidin-2-yl]-1-pentanol (14.13 g, yield: 72.5%).

NMR(CDCl₃) δ: 0.03(3H,s),0.05(3H,s),0.83(9H,s),1.43(9H,s),1.5-2.0 (6H,m),2.35(1H,m),3.03-3.23(2H,m),3.39(1H,m),3.55- 3.89(3H,m),4.17(1H,m),4.35(1H,m),4.70(1H,br),5.24 (2H,m),7.52(2H,br d,J=8Hz),8.24(2H,d,J=8Hz) ##STR1427##

To a solution of the compound obtained by the above reaction (1.56 g, 2.6 mmol) and triethylamine (0.37 ml, 2,6 mmol) in tetrahydrofuran (15 ml), methanesulfonyl chloride (0.21 ml, 2.6 mmol) was dropwise added under ice-cooling with stirring. After the mixture was stirred at the same temperature for 1 hour, the solvent was distilled off under reduced pressure. Trifluoroacetic acid (3.5 ml) was added to a solution of the residue in methylene chloride (10 ml) under ice-cooling with stirring, and the mixture was stirred at the same temperature for 1 hour. To a solution of the residue, which was obtained by evaporating the solvent and trifluoroacetic acid under reduced pressure. In tetrahydrofuran (20 ml), triethylamine (3 ml) was added, and the mixture was stirred overnight at room temperature. Di-tert-butyl dicarbonate (599 mg, 2.7 mmol ) was added to the reaction solution at room temperature with stirring. After the mixture was stirred at the same temperature for 1 four, the solvent was distilled off under reduced pressure, and ethyl acetate was added thereto. The organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was subjected to silica gel column chromatography (Wakogel™ C-300, heptane-ethyl acetate 2:1) to obtain (2S,4R)-2-(N-tert-butoxycarbonylpiperidin-4-yl)-4-tert-butyldimethylsiloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (1.05 g, yield: 69.9%).

NMR(CDCl₃) δ: 0.03(3H,s),0.05(3H,s),0.84(9H,s),0.96-1.30(1H,m), 1.35-1.62(1H,m),1.44(9H,s),1.74-1.90(2H,m),2.14(1H, m),2.18-2.76(1H,m),3.32(1H,m),3.61(1H,m),3.96-4.28 (3H,m),3.34(1H,m),5.24(2H,m),7.52(2H,d,J=8Hz),8.23 (1H,d,J=8Hz) ##STR1428##

To a solution of the compound obtained by the above reaction (1.0 g, 1.77 mmol) in tetrahydrofuran (10 ml), a solution of 1M tetrabutylammonium fluoride-tetrahydrofuran (2 ml, 2 mmol) was added under ice-cooling with stirring. The mixture was stirred at the same temperature for 40 minutes, and a saturated ammonium chloride aqueous solution (5 ml) was added thereto. After ethyl acetate (10 ml) was added thereto, the organic layer was separated, washed sequentially with water and a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. To a solution of the residue and triethylamine (0.25 ml, 1.78 mmol) in tetrahydrofuran (10 ml), methanesulfonyl chloride (0.14 ml, 1.8 mmol) was dropwise added under ice-cooling with stirring. After the mixture was stirred at the same temperature for 1 hour, ethyl acetate (10 ml) and water (10 ml) were added thereto. The organic layer was washed with a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, heptane-ethyl acetate 1:2) to obtain (2S,4R)-2-[N-(tert-butoxycarbonyl)piperidin-4-yl]-4-methanesulfonyloxy-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (852 mg, yield: 91.3%).

NMR(CDCl₃) δ: 1.0-1.3(2H,m),1.4-1.7(2H,m),1.44(9H,s),1.9-2.4(3H, m),1.45-1.76(2H,m),3.02(3H,s),3.45(1H,m),4.04-4.30 (4H,m),5.14-5.36(3H,m),7.54(2H,d,J=8Hz),8.25(2H,d, J=8Hz) ##STR1429##

To a solution of trityl mercaptan (4.21 g, 15.2 mmol) in N,N-dimethylformamide (50 ml), 60% sodium hydride (559 mg, 14 mmol) was added under ice-cooling with stirring, and the mixture was stirred at the same temperature for 20 minutes. A solution of the compound obtained by the above reaction (6.70 g, 12.7 mmol) in N,N-dimethylformamide (40 ml) was dropwise added to the reaction solution under ice-cooling with stirring. The mixture was brought to room temperature and stirred for 16 hours, then ethyl acetate (100 ml) and water (100 ml) were added to the reaction solution. The organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. The obtained residue was subjected to silica gel column chromatography (Wakogel™ C-300, heptane-ethyl acetate 2:1) to obtain (2S,4S]-2[N-(tert-butoxycarbonyl)piperidin-4-yl]-N-(p-nitrobenzyloxycarbonyl)-4-(tritylthio)pyrrolidine (7.34 yield: 81.6%).

NMR(CDCl₃) δ: 0.85-1.7(6H,m),1.44(9H,s),2.04(1H,m),2.46-2.76(4H, m),2.94(1H,m),3.68(1H,m),4.0-4.24(2H,m),5.0-5.28 (2H,m),7.1-7.6(17H,m),8.27(2H,d,J=8Hz) ##STR1430##

To a solution of the compound obtained by the above reaction (1.41 g, 2 mmol) in methylene chloride (2 ml), trifluoroacetic acid (1.5 ml) was added under ice-cooling with stirring. After the mixture was stirred at the same temperature for 1 hour, the solvent was distilled off under reduced pressure. To the obtained residue, ethyl acetate (10 ml) was added, and the mixture was washed sequentially with a saturated sodium hydrogencarbonate aqueous solution and a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. After 37% formalin (0.75 ml) was added to a solution of the residue in acetonitrile (8 ml) at room temperature, acetic acid (0.8 ml) was added thereto, and further sodium cyanoborohydride (377 mg) was added thereto. The mixture was stirred for 30 minutes, and the solvent was distilled off under reduced pressure. Ethyl acetate (10 ml) was added to the residue, washed sequentially with a saturated sodium hydrogencarbonate aqueous solution and a saturated sodium chloride aqueous solution, dried over anhydrous PG,383 sodium sulfate, and then the solvent was distilled off under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, methylene chloride-methanol 20:1) to obtain (2S,4S)-2-(N-methylpiperidin-4-yl)-N-(p-nitrobenzyloxycarbonyl)-4-(tritylthio)pyrrolidine (983 mg, yield: 79.0%).

NMR(CDCl₃) δ: 1.15-2.2(11H,m),2.28(3H,s),2.67(1H,m),2.87-3.0(2H, m),3.67(1H,m),5.0-5.24(2H,m),7.1-7.55(17H,m),8.28 (2H,d,J=8Hz) ##STR1431##

To a solution of the compound obtained by the above reaction (3.63 g, 5.8 mmol) in methylene chloride (10 ml trifluoroacetic acid (10 ml) was added under ice-cooling with stirring, and then triethylsilane (1.27 ml) was dropwise added thereto. After the mixture was stirred at the same temperature for 45 minutes, the solvent was distilled off under reduced pressure. The obtained residue was subjected to silica gel column chromatography (Wakeogel™ C300, methylene chloride-methanol 20:1→5:1) to obtain the above identified compound (2.30 g, yield: 80.6%).

NMR(CDCl₃) δ: 1.5-2.05(6H,m),2.25-2.88(3H,m),2.78(3H,s),2.98(1H, m),3.16(1H,m),3.52-3.72(2H,m),3.98(1H,m),4.14(1H, m),5.22(2H,br s),7.53(2H,d,J=8Hz),8.26(2H,d,J=8Hz)

REFERENCE EXAMPLE 38 (2S,4S)-4-mercapto-2-(N-methylpiperidin-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine hydrochloride ##STR1432##

To sodium metal (6.90 g, 0.30 mol), absolute ethanol (300 ml) was gradually added to prepare sodium ethoxide, and a solution of diethyl malonate (151.9 ml, 1.0 mol) in ethanol (500 ml) was dropwise added thereto at room temperature. A solution of (E)-3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-acrylic acid ethyl ester (400 g, 1.0 mol, compound of Reference Example 1-4) in ethanol (1500 ml) was further added thereto at the same temperature, and stirred overnight. After acetic acid (17.2 ml, 0.30 mol) was added thereto under cooling with ice, the reaction solution was concentrated under reduced pressure to obtain triethyl 2-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-1,1,3-propanetricarboxylate (558.3 g) as a crude oil.

To a solution of the compound obtained by the above reaction in dimethyl sulfoxide (2.8 l), lithium chloride (42.2 g, 1.0 mol) and water (35.9 ml, 2.0 mol) were added, and the mixture was stirred under heating at a temperature of from 143° to 160° C. for 1 hour (1 hour after gas evolving). The reaction solution was cooled, then extracted with ethyl acetate and washed with a sodium chloride aqueous solution. The organic layer was dried over anhydrous sodium sulfate and concentrated to obtain diethyl 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]glutarate (423.3 g) as a crude oil. ##STR1433##

To a solution of the compound obtained by the above reaction (447.8 g, 0.918 mol) in methanol (2.2 l), a 2N sodium hydroxide aqueous solution (1000 ml, 2.0 mol) was added at room temperature with stirring, and the mixture was stirred at the same temperature for 1.5 hours. The reaction solution was concentrated under reduced pressure, and ethyl acetate and dilute hydrochloric acid were added thereto. After the reaction solution was extractd with ethyl acetate, washed with dilute hydrochloric acid, the organic layer was dried and concentrated under reduced pressure. To the obtained oil, isopropyl ether (450 ml)/heptane (60 ml) was added, and the mixture was stirred overnight at 5° C. The precipitated crystal was collected by filtration to obtain 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyl-dimethylsiloxypyrrolidin-2-yl]glutaric acid (225.7 g, 56.9%) as first crystals. Second crystals were also collected in the same manner as above-mentioned to obtain the compound (18.9 g, 4.7%) (total 244.6 g, total yield: 61.7%).

IR(KBr)cm⁻¹ : 3600-3300, 2950, 2930, 2700-2400, 1730, 1680, 1650

NMR(CDCl₃) δ: 0.05(6H,s),0.S5(9H,s),1.47(9H,s),1.96-2.00(2H,m), 2.02-2.54(4H,m),3.20(1H,m),3.61(1H,m),3.93(1H,m), 4.30(1H,m) ##STR1434##

The compound obtained by the above reaction (225.7 g, 0.523 mol) was dissolved in tetrahydrofuran (3.3 l), and a borane-dimethyl sulfide complex (128.9 ml, 1.36 mol) was dropwise added thereto in a nitrogen stream under cooling with ice. After the mixture was stirred at room temperature for 2 hours, water (200 ml) and a saturated sodium hydrogencarbonate aqueous solution (100 ml) were added thereto. The mixture was extracted with ethyl acetate, and washed with a sodium chloride aqueous solution. The organic layer was dried and concentrated under reduced pressure, and then the obtained residue was subjected to silica gel column chromatography (Wakogel™ C-300, 3.5 E, heptane-ethyl acetate 3:1) to obtain 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyl-dimethylsiloxypyrrolidin-2-yl]-1,5-pentanediol (117.9 g, yield: 55.9%).

IR(KBr)cm⁻¹ : 3700-3100, 2940, 2855, 1700, 1680

NMR(CDCl₃) δ: 0.04(6H,s),0.86(9H,s),1.45(9H,s),1.66-1.94(2H,m), 2.38(1H,m),3.24(1H,m),3.40-3.82(4H,m),3.99(1H,m), 4.09(1H,m) ##STR1435##

To a solution of the compound obtained by the above reaction (251 g, 0.629 mol) in tetrahydrofuran (2 l), methanesulfonyl chloride (107 ml, 1.38 mol) and triethylamine (192 ml, 1.38 mmol) were dropwise added under cooling with ice, and the mixture was stirred at room temperature for 30 minutes. After the reaction mixture was filtered, the filtrate and the washings were put together, and the mixture was concentrated under reduced pressure. Ethyl acetate (1 l) was added to the residue, and washed sequentially with dilute hydrochloric acid (200 ml), a saturated sodium hydrogencarbonate aqueous solution (200 ml×2) and a saturated sodium chloride aqueous solution (200 ml). The reaction solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(1,5-dimethanesulfonyloxypent-3-yl)pyrrolidine (302 g, yield: 86%) as a crude product. ##STR1436##

To a solution of the compound obtained by the above reaction (302 g, 0.544 mol) in methanol (700 ml), a 40% methylamine/methanol solution (400 ml) was added under cooling with ice, and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure, and then to the obtained residue, ethyl acetate (1 l) and a 2N sodium hydroxide aqueous solution (200 ml) were added. The organic layer was washed with the mixture of a 5N sodium hydroxide aqueous solution (20 ml) and a saturated sodium chloride aqueous solution (200 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, chloroform-methanol 50:1→1:5) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-methylpiperidin-4-yl)pyrrolidine (100 g, yield: 46%).

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.46(9H,s),2.29(3H,s),2.76-3.00(3H,m),3.34(1H,dd,J=12,4Hz),3.50(1H,m),3.87 (1H,m),4.30(1H,m) ##STR1437##

3N Hydrogen chloride-methanol was added to the compound obtained by the above reaction (100 C, 0.25 mol) at room temperature, and the mixture was stirred overnight at the same temperature. After the reaction mixture was concentrated under reduced pressure, ethanol (200 ml) was added to the residue, and then concentrated under reduced pressure. The same procedure was carried out three times, and ethanol (100 ml) was added to the precipitates. The precipitates were collected by filtration in a nitrogen stream, washed with ethanol, and then dried in a nitrogen stream to obtain (2S,4R)-4-hydroxy-2-(N-methylpiperidin-4-yl)pyrrolidine dihydrochloride (41 g, yield: 63%).

NMR(D₂ O) δ: 1.38-1.74(2H,m),1.74-2.06(4H,m),2.17(1H,m)2.81(3H, s),2.88-3.08(2H,m),3.25(1H,m),3.36-3.68(4H,m),4.51 (1H,m) ##STR1438##

To the compound obtained by above reaction (36 g, 0.14 mol), absolute N,N-dimethylformamide (360 ml) and triethylamine (65 ml, 0.462 ml) were added at room temperature, and the mixture was stirred at the same temperature for 15 minutes. After a solution of p-nitrobenzyloxycarbonyl chloride (33.4 g, 0.155 mol) in N,N-dimethylformamide (50 ml) was added thereto at 0° C., the mixture was stirred at the same temperature for 1 hour. p-Nitrobenzyloxycarbonyl chloride (3.3 g, 0.015 mol) was added thereto, and the mixture was stirred for 10 minutes. After the reaction mixture was filtered, the filtrate and the washings were put together, and the mixture was concentrated under reduced pressure. Ethyl acetate (100 ml) was added to the obtained oily substance, and then extracted with 3N hydrochloric acid (100 ml, 20 ml×2). After the aqueous layer was washed with ethyl acetate (100 ml), a 5N sodium hydroxide aqueous solution (250 was added thereto, and the mixture was extracted with ethyl acetate (300 ml, 100 ml ×2). The organic layer was washed with a mixture of a 5N sodium hydroxide aqueous solution (5 ml) and a saturated sodium chloride aqueous solution (100 ml), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain (2S,4R)-4-hydroxy-2-(N-methylpiperidin-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (30.2 g, crude yield: 59%).

NMR(CDCl₃) δ: 1.10-2.10(11H,m),2.14(3H,S),2.90(2H,m),S.41(1H,m), 3.69(1H,m),4.08(1H,m),4.42(1H,m),5.24(2H,m),7.52 (2H,d,J=8Hz),8.24(2H,d,J=8Hz) ##STR1439##

To a solution of the compound obtained by the above reaction (16.85 g, 45.7 mmol) in methylene chloride (340 ml), triethylamine (6.37 ml, 45.7 mmol) was added at 0° C., and methanesulfonyl chloride (3.54 ml, 45 7 mmol) was dropwise added thereto over a period of 7 minutes. The mixture was stirred at the same temperature for 30 minutes, and methylene chloride (340 ml) was added thereto. The reaction mixture was washed sequentially with a mixture of ice (150 ml) and a saturated sodium hydrogencarbonate aqueous solution (150 ml), and a saturated sodium chloride aqueous solution (200 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain (2S,4R)-4-methanesulfonyloxy-2-(N-methylpiperidin-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (18.87 g, yield: 92%).

NMR(CDCl₃) δ: 1.22-1.72(4H,m),2.28(3H,s),2.82-3.00(2H,m),3.02(3H, s),3.51(! H,m),4.02-4.22(2H,m),4.01-4.22(2H,m),7.54 (2H,d,J=8Hz),8.22(2H,d,J=8Hz) ##STR1440##

A solution of the compound obtained by the above reaction (2.1 g, 4.7 mmol), potassium thioacetate (700 mg, 6 mmol) and sodium iodide (785 mg, 5.2 mmol) in N,N-dimethylformamide (20 ml) was stirred in a nitrogen stream at 60° C. for 16 hours. To the reaction mixture, ethyl acetate (50 ml) and water (50 were added. The organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure. The obtained residue was subjected to silica gel column chromatography (Wakogel™ C-300, methylene chloride-methanol 1:5→1:20) to obtain (2S,4S)-4-acetylthio-2-(N-methyl-piperidin-4-yl)-N-(p-nitrobenzyloxycarbonyl)pyrrolidine (1.56 g, yield: 77.3%).

NMR(CDCl₃) δ: 1.2-2.2(8H,m),2.34(6H,s),2.66(1H,m),2.88-3.08(3H, m),3.78(1H,m),S.98(1H,m),4.26(1H,m),5.23(2H,br s), 7.52(2H,d,J=8Hz),8.24(2H,d,J=8Hz) ##STR1441##

To a solution of the compound obtained by the above reaction (3.03 g, 7.2 mmol) in methanol (30 ml), a 1N sodium methoxide-methanol solution (3.6 ml, 3.6 mmol) was added, and the mixture was stirred in a nitrogen stream at room temperature for 30 minutes. After a 3N hydrogen chloride-methanol solution (7 ml) was added to the reaction mixture under ice-cooling with stirring, the solvent was distilled off under reduced pressure. The obtained residue was subjected to silica gel column (Wakogel™ C-300, chromatography methylene chloride-methanol 20:1→5:1) to obtain the above identified compound (2.76 g, yield: 92.3%).

NMR(CDCl₃) δ: 1.54-2.86(9H,m),2.78(3H,s),2.86-3.28(2H,m),3.44- 3.68(2H,m),3.92-4.24(2H,m),5.23(2H,br s),7.54(2H, d,J=8Hz),8.27(2H,d,J=8Hz)

REFERENCE EXAMPLE 39 (2S,4R)-4-hydroxy-2-(N-methylpiperidin-4-yl)pyrrolidine dihydrochloride ##STR1442##

To a suspension of 60% sodium hydride (10 g, 250 mmol) in tetrahydrofuran (700 ml), ethyl cyanoacetate (26.6 ml, 250 mmol) and tetra-n-butylammonium bromide (8.06 g, 25 mmol) were added in a nitrogen stream under ice-cooling with stirring, and the mixture was stirred for 30 minutes. A solution of (E)-3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-acrylic acid ethyl ester (100 g, 250 mmol, compound of Reference Example in tetrahydrofuran (300 ml was dropwise added to the reaction solution under ice-cooling with stirring over a period of 30 minutes. The reaction solution was stirred at 50° C. for 20 hours neutralized with acetic acid, and then poured into ethyl acetate (1000 ml) and a hydrochloric acid aqueous solution (pH6). The organic layer was washed with a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain a brown oily substance (127.43 g, crude yield: 99.5%) containing diethyl 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-2-cyanoglutarate.

NMR(CDCl₃) δ: 0.04(6H,s),0.84(9H,s),1.48(9H,s),1.12-1.34(6H,m), 1.70-2.12(2H,m),2.30-2.66(2H,m),3.04-3.84(4H,m), 4.02-4.38(6H,m) ##STR1443##

To a solution of the compound obtained by the above reaction (127.14 g, 250 mmol) in dimethyl sulfoxide (700 ml), sodium chloride (14.6 g, 250 mmol) and water (9 ml 500 mmol) were added, and the mixture was stirred at 144° C. for 3 hours. After the reaction mixture was poured into a mixed solution of ice water (700 ml) and ethyl acetate (700 ml), the organic layer was washed sequentially with water and a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfane, and then concentrated under reduced pressure to obtain a brown oily substance (101.2 g, crude yield: 92%) containing ethyl 3-[(2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin-2-yl]-4-cyanobutyrate.

NMR(CDCl₃ ) δ: 0.06(6H,s),0.86(9H,s),1.28(3H,t,J=7Hz),1.46(9H,s), 1.64-2.08(2H,m),2.30-2.70(4H,m),2.79(1H,m),3-20(1H, m),3.54(1H,m),4.18(2H,q,J=7Hz),4.11(1H,m),4-34(1H, br s) ##STR1444##

To a solution of the compound obtained by the above reaction (50 g) in methanol (800 ml), Raney Ni (about 50 g) was added, and the mixture was stirred in a hydrogen stream (3 arm.) at room temperature for 22 hours. The catalyst was filtered off from the reaction mixture, and the filtrate was concentrated under reduced pressure obtain a brown oily substance (42.3 g, crude yield: 93.5%) containing (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-piperidon-4-yl)pyrrolidine.

NMR(CDCl₃) δ: 0.05(6H,s),0.86(9H,s),1.45(9H,s),1.71-1.97(4H,m), 2.21-2.47(2H,m),3.11-3.55(5H,m),3.99(1H,m),4-31(1H, m),5.91(1H,br s) ##STR1445##

To a solution of the compound obtained by the above reaction (42.3 g) in tetrahydrofuran (400 ml), 60% sodium hydride (4.5 g, 111 mmol) was added in a nitrogen stream under ice-cooling with stirring. The mixture was stirred for 20 minutes, then methyl iodide (13.2 ml, 212 mmol) was added thereto and the mixture was stirred in a nitrogen stream at room temperature for 18.5 hours. After a saturated ammonium chloride aqueous solution was added to the reaction solution under cooling with ice, the mixture was poured into a mixed solution of ethyl acetate (400 ml) and water (400 ml). The organic layer was washed sequentially with a saturated sodium sulfite aqueous solution and a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain a yellow oily substance (39.3 g, crude yield: 89.8%) containing (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-methyl-2-piperidon-4-yl)pyrrolidine

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.46(9H,s),1.62-2.08(4H,m), 2.28-2.54(2H,m),2.94(3H,s),3.04-3.73(5H,m),4-02(1H, m),4.30(1H,m) ##STR1446##

To a solution of the compound obtained by the above reaction (5.98 g) in tetrahydrofuran (60 ml), a borane-dimethyl sulfide complex (4.13 ml, 43.5 mmol) was added in a nitrogen stream under ice-cooling with stirring, and the mixture was refluxed in a nitrogen stream for 2 hours. After methanol (30 ml) was added to the reaction solution under ice-cooling with stirring, the mixture was concentrated under reduced pressure. The oily residue was poured into a mixed solution of ethyl acetate and water, and the organic layer was washed with a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain a yellow oily substance (5.80 g, crude yield: 100.4%) containing (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-methylpiperidin-4-yl)pyrrolidine-borane.

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.46(9H,s),1.62-1.98(6H,m), 2.56 and 2.62(3H,s),2.82-3.23(5H,m),3.40-3.70(2H, m),4.00(1H,m),4.30(1H,m) ##STR1447##

To a 3N hydrogen chloride-methanol solution, compound obtained by the above reaction (5.80 g) was added, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure to obtain white crystals (3.85 g, crude yield: 102.7%) containing the above identified compound.

NMR(D₂ O) δ: 1.38-1.74(2H,m),1.74-2.06(4H,m),2.14(1H,m),2.8i(3H, s),2.88-3.08(2H,m),3.25(1H,m),B.36-3.68(4H,m),4.61 (1H,m)

REFERENCE EXAMPLE 40 (2S,4S)-2-(N-ethylpyrrolidin-3-yl)-4-mercaptopyrrolidine dihydrochloride ##STR1448##

To a solution of (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-pyrrolidon-4-yl)pyrrolidine (38.5 g, 0.1 mol, the compound of Reference Example 1-6) in tetrahydrofuran (200 ml) was added 60% sodium hydride in oil (4.2 g, 0.1 mol) under ice-cooling, and the mixture was stirred for 1 hour. To the mixture was added ethyl iodide (8 rag, 0.1 mol), and after stirring at room temperature overnight, the reaction mixture was concentrated. The residue was extracted with ethyl acetate (500 ml), and the mixture was washed with water (200 ml) followed by a saturated sodium chloride aqueous solution. After the extract was dried over anhydrous magnesium sulfate, the solvent was removed in vacuo, and the residue was purified by silica gel column chromatography (Wakogel™ C-300, chloroform) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-ethyl-2-pyrrolidon-4-yl)pyrrolidine (40.3 g, yield: 97.7%).

IR(KBr)cm⁻¹ : 3450, 2920, 1690, 1390, 1360, 1250

NMR(CDCl₃) δ: 0.06(6H,s),0 88(9H,s),1 15(3H,t,J=9Hz),1.48(9H,s), 1.8-2.1(2H,m),2.2-2.6(2H,m),3.1-3.5(6H,m),3.6(1H, m),4.12(1H,m),4.35(1H,m) ##STR1449##

To solution a of the compound (40 g, 96.9 mmol) obtained by the above reaction in tetrahydrofuran (200 ml) was added borane-dimethylsulfide complex (23.01 ml, 242 mmol) under cooling with ice, and the mixture was refluxed for 30 minutes. After cooling the reaction mixture, methanol (100 ml) was added thereto, and then stirred for 10 minutes at room temperature. After the solvent was removed in vacuo, the residue was extracted with ethyl acetate (500 ml), and the extract was washed with a saturated sodium chloride aqueous solution (200 ml×2). The extract was dried over anhydrous sodium sulfate, and concentrated to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-ethylpyrrolidin-3-yl)pyrrolidine-borane complex (35.9 g, yield: 89.7%).

IR(KBr)cm⁻¹ : 3450, 2920, 2370, 1690, 1460, 1390 NMR(CDCl₃) δ: 0.06(6H,s),0.88(9H,s),1.15(3H,=,J=9Hz),1.48(9H,s), 1.8-2.1(2H,m),2.5-3.0(4H,m),8.2(2H,m),3.49(1H,m), 4.1(1H,m),4.32(1H,m) ##STR1450##

To solution of a the compound obtained by the above reaction (35.8 g, 86.8 mmol) in tetrahydrofuran (200 ml) was added 1 M tetrabutylammonium fluoride in tetrahydrofuran (91.1 ml) under cooling with ice, and the mixture was stirred for 1 hour at room temperature. To the reaction mixture was added ethyl acetate (500 ml), and the organic layer was washed with water (200 ml) followed by a saturated sodium chloride aqueous solution (100 ml). The solvent was dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography (Wakogel™ C-300, 5% methanol-chloroform) to obtain (2S, 4R)-N-tert-butoxycarbonyl-2-(N-ethylpyrrolidin-3-yl)-4-hydroxypyrrolidine-borane complex (44.1 g).

IR(KBr)cm⁻¹ : 3300, 2970, 2370, 1690, 1460, 1390, 1250

NMR(CDCl₃) δ: 1.32(3H,t,J=9Hz),1.48(9H,s),1.8-2.3(2H,m),2.7-3.0 (6H,m),3.2-3.3(2H,m),3.6(1H,m),4.2(1H,m),4.5(1H,m) ##STR1451##

To a solution of the compound obtained by the above reaction (44 g, 86.8 mmol) in methylene chloride (300 ml) was added triethylamine (13.3 ml, 95.5 mmol) and methanesulfonyl chloride (7.4 ml, 95.5 mmol) under cooling with ice, and the mixture was stirred for 1 hour at the same temperature. To the reaction mixture was added methylene chloride (200 ml), and the organic layer was washed with water (200 ml) followed by a saturated sodium chloride aqueous solution (100 ml). The solvent was dried over anhydrous magnesium sulfate and concentrated to obtain (2S,4R)-N-tert-butoxycarbonyl-2-(N-ethylpyrrolidin-3-yl)-4-methanesulfonyloxypyrrolidine-borane complex (30.39 g, yield: 80.6%). IR(KBr)cm⁻¹ : 450, 2970, 2360, 1690, 1450, 1390, 1360

NMR(CDCl₃) δ: 1.35(3H,t,J=9Hz),1.48(9H,s),1.8-2.3(2H,m),2.3-3.0 (8H,m),3.06(3H,s),3.25(2H,m),3.4(1H,m),4.2(1H,m), 5.24(1H,m) ##STR1452##

To a suspension of anhydrous potassium carbonate (11.6 g, 84 mmol) in N,N-dimethylformamide (300 ml) was added thioacetic acid (6.0 ml, 84 mmol) under cooling with ice, and the mixture was stirred for 30 minutes under a nitrogen atmosphere. To the mixture was added sodium iodide (11.5 g, 77 mmol) and a solution of the compound obtained by the above reaction (30 g, 70 mmol) in N,N-dimethylformamide (100 ml), and the mixture was stirred overnight at 60°-70° C. To the reaction mixture was added ethyl acetate (1 l), and the mixture was washed with water (500 ml×3 ) followed by a saturated sodium chloride aqueous solution (100 ml×2). The extract was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column chromatography (Wakogel™ C-300, chloroform) to obtain (2S,4S)-4-acetylthio-N-tert-butoxycarbonyl-2-(N-ethylpyrrolidin-3-yl)pyrrolidine-borane complex (17.3 g, yield: 59.6%). IR(KBr)cm⁻¹ : 3420, 2970, 2360, 1700, 1690, 1390, 1360

NMR(CDCl₃) δ: 1.3(3H,t,J=9Hz),1.48(9H,s),1.8-2.3(2H,m),2.5(2H,m), 2.6-3.1(6H,m),3.2(2H,m),3.8(1H,m),4.1(2H,m) ##STR1453##

The compound obtained by the above reaction (17 g, 41 mmol) was dissolved in 1.18 N hydrochloric acid-methanol (104 ml), and the solution was stirred an temperature for 1 hour followed by at 60°-70° C. for 4 hours. The excess hydrochloric acid-methanol was removed in vacuo, and ethanol (30 ml) was added to the residue. The resulting crystal was collected by filtration, washed with cold ethanol, and dried to obtain the above identified compound (7.1 g, yield: 63.6%). The mother liquor was concentrated to obtain the second crystal (2.8 g, yield: 25.1%)

IR(KBr)cm⁻¹ : 3400, 2930, 2690, 1450, 1410, 1115

NMR(DMSO-d₆) δ: 1.23(3H,t,J=9Hz),1.6(1H,m),1.8-2.4(2H,m),2.52(1H, m),2.95(1H,m),2.15(4H,m),2.2-2.7(7H,m)

mp: 178° C.

REFERENCE EXAMPLE 41 (2S,4S)-2-(azetidin-3-yl)-4-mercapto-N-(p-nitrobenzyloxycarbonyl) pyrrolidine ##STR1454##

To a solution of (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-nitroylnyl)pyrrolidine (7.02 g, 18.8 mmol, compound of Reference Example 31-2) in chloroform (80 ml), silica gel (Wakogel™ C-300, 10 g) and isopropanol (10 ml) were added. To this solution was added sodium borohydride (3.40 g, 90 mmol) 0° C. The mixture was stirred at room temperature for 3 hours, then cooled in an ice-bath, water (30 ml) d 3 N hydrochloric acid were added thereto to adjust to pH 4, then brought to pH 10 with 3 N sodium hydroxide. The mixture was filtered, then extracted with chloroform (200 ml). The organic layer was washed sequentially with a saturated ammonium chloride aqueous solution, a saturated sodium hydrogencarbonate aqueous solution and a saturated sodium chloride aqueous solution, dried ever anhydrous sodium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 8:1→6:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-nitroethyl)pyrrolidine (4.68 g, yield: 67%).

NMR(CDCl₃) δ: 0.04(6H,s),0.84(9H,s),1.44(9H,s),1.64(1H,m),2.04 (1H,m),2.14-2.44(2H,m),3.20-3.60(2H,m),4.04(1H,m), 4.32(1H,m),4.48(2H,m) ##STR1455##

To 60% sodium hydride (2.56 g, 64 mmol), tert-butylalcohol (50 ml) was added in a nitrogen stream at room temperature and the mixture was stirred for 20 minutes. To this mixture, a solution of (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(2-nitroethyl)pyrrolidine (11.8 g, 32 mmol) in tert-butylalcohol (30 ml) was added at room temperature. After stirring for 20 minutes, the mixture was added to precooled heptane (1 l) in an ice-bath and further stirred for to minutes at 0° C.. To the reaction mixture, a solution of potassium permanganate (5.06 g, 32 mmol) in water (100 ml) and a solution of boric acid (3.94 g, 64 mmol) in water (50 ml) were added, and the mixture stirred for minutes. To the reaction mixture, a solution of sodium thiosulfate pentahydrate (15.88 g, 64 mmol) in water ml) and 1 N sulfuric acid (64 ml) were added. After stirring for 10 minutes, the reaction mixture was filtered with celite and washed with heptane (200 ml). The filtrate and washings were combined, and the organic layer was washed sequentially with a saturated sodium hydrogencarbonate aqueous solution (200 ml) and a saturated sodium chloride aqueous solution (200 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained oily residue was subjected to silica gel column chromatography (Wakogel™ C-300, heptane-ethyl acetate 5:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(formylmethyl)pyrrolidine (7.0 g, yield: 66%).

NMR(CDCl₃) δ: 0.04(6H,s),0.86(9H,s),1.44(9H,s),1.72(1H,m),2.15 (1H,m),2.52(1H,m),2.90(1H,m),3.38(2H,m),4.33(2H,m), 9.78 (1H, t,J=2Hz) ##STR1456##

To a solution of the compound obtained by the above reaction (2.92 g, 8.81 mmol) in dry methylene chloride (30 ml), N,N-dimethylmethyleneammonium chloride (1.65 q, 17.6 mmol) and triethylamine (2.8 ml, 20 mmol) were added at room temperature and the reaction mixture was stirred overnight at the same temperature. After addition of chloroform (100 ml), the mixture was washed with water and a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 3:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(1-formylylnyl)pyrrolidine (2.54 g, yield: 81%).

NMR(CDCl₃) δ: 0.07(6H,s),0.88(9H,s),1.28(4.5H,br s),1.47(4.5H, br s),1.78(1H,m),2.22(1H,m),3.52(2H,m),4.32(1H,m), 4.76(1H,m),6.06(1H,br s),6.28(1H,br s),9.62(1H,s) ##STR1457##

To a solution of the compound obtained by the above reaction (2.45 g, 6.9 mmol) in methanol (10 ml), a solution of cerium trichloride hexahydrate (2.50 G, 7.0 mmol) in methanol (10 ml) was added. To the mixture was added at 0° C. sodium borohydride (270 mg, 7.1 mmol), and the mixture stirred for 10 minutes at room temperature. After addition of water (15 ml) and ethyl acetate (80 ml the organic layer was washed sequentially with dilute hydrochloric acid, water, a saturated sodium hydrogen-carbonate aqueous solution and a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure obtain crude (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyl-dimethylsiloxy-2-[1-(hydroxymethyl)ylnyl]pyrrolidine (2.4 g, yield: 97%).

NMR(CDCl₃) δ: 0.06(6H,s),0.88(9H,s),1.42(9H,s),1.94(1H,m),2.08 (1H,m),3.26(2H,br s),4.01(2H,br s),4.38(1H,m),4.51 (1H,m),5.02(1H,br s),5.12(1H,br s) ##STR1458##

To a solution of the compound obtained by the above reaction (4.09 g, 11.4 mmol) in dry methylene chloride (40 ml), 3,4-dihydropyran (2.1 ml, 23 mmol) and pyridinium p-toluenesulfonate (300 mg, 1.2 mmol) were added at room temperature. After stirring for 6 hours at the same temperature, the reaction mixture was washed with water (20 ml×2) and a saturated sodium chloride aqueous solution (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyl-dimethylsiloxy-2-[1-(tetrahydropyranyloxymethyl)vinyl]pyrrolidine (4.7 g, yield: 93%).

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.41(9H,s),3.50(3H,m),3.74-4.54(5H,m),4.64(1H,m),5.01(1H,br s),5.10(1H,br s) ##STR1459##

To a solution of the compound obtained by the above reaction (550 mg, 1.25 mmol) in dry tetrahydrofuran (5 ml), a solution of 9-borabicyclo[3.3.1]nonane (340 mg, 1.4 mmol) in dry tetrahydrofuran (5 ml) was added at room temperature, and the mixture stirred overnight at the same temperature. To this solution, sodium perborate tetrahydrate (770 mg, 5 mmol) and water (5 ml) were added tinder cooling with ice, and the mixture shirred for 7 hours at room temperature. To the reaction mixture, ethyl acetate (100 ml) was added, then the mixture washed with water (50 ml) and a saturated sodium chloride aqueous solution (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained oily residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 3:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[2-hydroxyl-1-(tetrahydropyranyloxymethyl)ethyl]pyrrolidine (542 mg, yield: 94%).

NMR(CDCl₃) δ: 0.04(6H,s),0.86(9H,s),4.34(1H,m),4.58(1H,m) ##STR1460##

The same procedure as in Reference example 32-3) was carried out by using the compound obtained by the above reaction to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[2-phthalimido-1-(tetrahydropyranyloxymethyl)ethyl] pyrrolidine (1.94 g, yield: 94.2%).

NMR(CDCl₃) δ: 0.06(6H,s),0.86(9H,s),1.20-1.70(15H,m),1.94-2.15 (2H,m),4.36(1H,m),4.48(1H,m),7.72(2H,m),7.84(2H,m) ##STR1461##

To a solution of the compound obtained by the above reaction (1.94 g, 3.3 mmol) in ethanol (20 ml), hydrazine monohydrate (0.48 ml, 9.9 mmol) was added and the reaction mixture was stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure and the residue was extracted with ethyl acetate (100 ml). The organic layer was washed sequentially with 2 N aqueous ammonia (×3) and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was dissolved in tetrahydrofuran (25 ml) , and to the resulting solution, triethylamine (0.58 ml, 4.17 mmol) and p-toluenesulfonyl chloride (630 mg, 3.3 mmol) were added under cooling with ice. The mixture was stirred for 1.5 hours at the same temperature and extracted with ethyl acetate (100 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 6:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[1-(tetrahydropyranyloxymethyl)-2-(p-toluenesulfonylamino)ethyl]pyrrolidine (1.38 g, yield: 68.1%).

NMR(CDCl₃) δ: 0.03(6H,s),0.85(9H,s),1.38-1.98(17H,m),2.43(3H,m), 4.27(1H,m),4.42-4.62(1H,m),7.30(2H,br d,J=8Hz),7.76 (2H,br d,J=8Hz) ##STR1462##

To a solution of the compound obtained by the above reaction (1.38 g, 2.25 mmol) in methanol (40 ml), pyridinium p-toluenesulfonate (60 mg, 0.24 mmol) was added and the reaction mixture was stirred for 4.5 hours at 45°-50 ° C. The resulting mixture was concentrated under reduced pressure and the residue was extracted with ethyl acetate (70 ml). The organic layer was washed with water and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 3:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[1-hydroxymethyl-2-(p-toluenesulfonyl-amino)ethyl]pyrrolidine (912 mg, yield: 76.6%).

NMR(CDCl₃) δ: 0.06(6H,s),0.87(9H,s),1.40-1.50(9H,m),1.75(1H,m), 2.00(1H,m),2.44(3H,m),7.30(2H,m),7.76(2H,m) ##STR1463##

To a solution of the compound obtained by the above reaction (912 mg, 1.73 mmol) in tetrahydrofuran (15 ml), triethylamine (0.288 ml, 2.07 mmol) and methanesulfonyl chloride (0.153 ml, 1.98 mmol) were added successively under cooling with ice and the mixture was stirred for 40 minutes at the same temperature. The reaction mixture was extracted with ethyl acetate (70 ml) and the organic layer was washed with water and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[1-(methanesulfonyloxymethyl)-2-(p-toluenesulfonylamino)ethyl]pyrrolidine (980 mg, yield: 93.6%).

NMR(CDCl₃) δ: 0.05(6H,s),0.85(9H,s),1.36-1.46(9H,m),1.80 (1H,m), 1.98(1H,m),2.44(3H,m),2.90-3.20(6H,m),3.60 1H,m), 4.08-4.35(4H,m),7.30(2H,m),7.74(2H,m) ##STR1464##

To a solution of the compound obtained by the above reaction (980 mg, 1.62 mmol) in N,N-dimethylformamide (80 ml), 60% sodium hydride (97 mg, 2.42 mmol) was added under nitrogen atmosphere under cooling with ice, and the resulting mixture was stirred for 15 minutes at the same temperature, then for 4 hours at room temperature. The reaction mixture was concentrated under reduced pressure and the residue was extracted with ethyl acetate (70 ml). The organic layer was wasted with water and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 8:1) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[N-(p-toluenesulfonyl)azetidin-3-yl]pyrrolidine (682 mg, yield: 82.7%).

NMR(CDCl₃) δ: 0.02(6H,s),0.82(9H,s),1.41(9H,s),1.58(1H,m),1.87 (1H,m),2.46(3H,s),2.76(1H,m),3.15(1H,m),3.42(2H,m), 3.73(3H,m),3.90(1H,m),4.22(1H,m),7.38(2H,d,J=8Hz), 7.73(2H,d,J=8Hz) ##STR1465##

To a solution of the compound obtained by the above reaction (682 mg, 1.34 mmol) in tetrahydrofuran (10 ml), liquid ammonia (ca. 20 ml) was added under nitrogen atmosphere at -78° C., and to the resulting solution was added sodium metal (140 mg, 6.1 mmol). The reaction mixture was stirred for 30 minutes at the same temperature, then the solvent was distilled off. To the residue in tetrahydrofuran (20 ml), triethylamine (0.186 ml, 1.34 mmol) and 4,6-dimethyl-2-(p-nitrobenzyloxycarbonylthio)pyrimidine (383 mg, 1.20 mmol) were added under cooling with ice. The reaction mixture was stirred for 2 hours at room temperature, then extracted with ethyl acetate (70 ml). The organic layer was washed sequentially with a 1N sodium hydroxide aqueous solution, water and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, hexane-ethyl acetate 5:1) no obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-[N-(p-nitrobenzyloxycarbonyl)azetidin-3-yl]pyrrolidine (283 mg, yield: 39.5%).

NMR(CDCl₃) δ: 0.06(6H,s),0.87(9H,s),1.46(9H,s),1.74(1H,m),2.02 (1H,m),2.96(1H,m),3.30(1H,m),3.42-3.86(2H,m),3.90-4.14(3H,m),4.15-4.38(2H,m),5.20(2H,s),7.53(2H,d,J=9Hz),8.23(2H,d,J=9Hz) ##STR1466##

The compound obtained by the above reaction (283 mg, 0.53 mmol) was dissolved in 0.65 N hydrogen chloride-methanol (5 ml) and the resulting mixture was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure and the residue was dissolved in dioxane (5 ml) and water (2.5 ml). The mixture was adjusted to pH 9 with a 1 N aqueous sodium hydroxide aqueous solution and 4,6-dimethyl-2-(p-nitrobenzyloxycarbonylthio)pyrimidine (170 mg, 0.53 mmol) was added thereto. The resulting mixture was stirred overnight at room temperature, then concentrated under reduced pressure. The residue was extracted with ethyl acetate (50 ml). The organic layer was washed sequentially with a 1 N sodium hydroxide aqueous solution, water and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 1% methanol-chloroform) to obtain (2S, 4R)-4-hydroxy-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)azetidin-3-yl]pyrrolidine (87 mg, yield: 32.9%).

NMR(CDCl3) δ: 1.86(1H,m),2.18(1H,m),3.02(1H,m),3.10-3.62(2H,m), 3.64-3.90(2H,m),3.94-4.20(2H,m),4.34(1H,m),4.50(1H, m),5.10-5.30(4H,m),7.51(4H,d,J=9Hz),8.22(4H,m) ##STR1467##

To a solution of the compound obtained by the above reaction (87 mg, 0.174 mmol) in tetrahydrofuran (5 triphenylphosphine (92 mg, 0.35 mmol) and diethyl azodicarboxylate (55 μl, 0.35 mmol) were added sequentially under nitrogen atmosphere under cooling with ice and the reaction mixture was stirred for 30 minutes at the same temperature. To the mixture, thioacetic acid (31 μl, 0.43 mmol) was added and the resulting mixture was further stirred for 2 hours under cooling with ice. The reaction mixture was extracted with ethyl acetate (50 ml) and the organic layer was washed with water and a saturated sodium chloride aqueous solution, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by preparative TLC (Merck silica gel 60 F₂₅₄, hexane-ethyl acetate 1:3) to obtain (2S, 4S)-4-acetylthio-N-(p-nitrobenzyloxycarbonyl)-2-[N-(p-nitrobenzyloxycarbonyl)azetidin-3-yl]pyrrolidine (78.9 mg, yield: 81.3%).

NMR(CDCl₃) δ: 1.65(1H,m),2.36(3H,s),2.58(1H,m),3.05(1H,m),3.23 (1H,m),3.66-4.30(7H,m),5.12-5.18(4H,m),7.52(4H,m), 8.23(4H,m) ##STR1468##

The same procedure as in Reference Example 32-10) was carried out by using the compound obtained by the above reaction (78.9 mg, 0.14 mmol) to obtain the above identified compound (73 mg, yield: 100% ).

REFERENCE EXAMPLE 42 (2S,4S)-4-mercapto-2-(N-methylazetidin-3-yl-N-(p-nitrobenzyloxycarbonyl)pyrrolidine hydrochloride ##STR1469##

To a solution of (2S,4R)-2-(azetidin-3-yl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidin (850 mg, 2.39 mmol) obtained by the reaction of Reference Example 41-12) in acetonitrile (10 ml) were added 37% formalin (0.75 ml) and acetic acid (0.8 ml) at room temperature. To the mixture, sodium cyanoborohydride (370 mg, 5.9 mmol) was further added and the resulting mixture was stirred for 30 minutes. The reaction mixture was concentrated under reduced pressure and the residue was extracted with ethyl acetate (50 ml). The organic layer was washed sequentially with a saturated sodium hydrogencarbonate aqueous solution and a saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 10% methanol-chloroform) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-methylazetidin-3-yl)pyrrolidine (620 mg, yield: 70%).

IR(KBr)cm⁻¹ : 1700, 1400, 1250, 1170 ##STR1470##

To a solution of (2S,4R)-2-(azetidin-3-yl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidine (850 mg, 2.39 mmol) obtained by the reaction of Reference Example 41-12) in acetonitrile (10 ml) were added 37% formalin (0.75 ml) and acetic acid (0.8 ml) at room temperature. To the mixture, sodium cyanoborohydride (370 ml, 5.9 mmol) was further added and the resulting mixture was stirred for 30 minutes. The reaction mixture was concentrated under reduced pressure and the residue was extracted with ethyl acetate (50 ml). The organic layer was washed sequentially with a saturated sodium hydrogencarbonate aqueous solution and saturated sodium chloride aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (Wakogel™ C-300, 10% methanol-chloroform) to obtain (2S,4R)-N-tert-butoxycarbony-1-4-tert-butyldimethylsiloxy-2-(N-methylazetidin-3-yl)pyrrolidine (620 ml, yield: 70%).

IR(KBr)cm⁻¹ : 1700, 1400, 1250, 1170 ##STR1471##

The same procedures as in Reference Example 38-6), 7), 8), 9), 10) were carried out by using the compound obtained by the above reaction (620 mg, 1.68 mmol) to obtain the above identified compound (195 mg).

IR(KBr)cm⁻¹ : 1700, 1520, 1350, 1110

REFERENCE EXAMPLE 43 (2S,4S)-2-(N-ethylazetidin-3-yl)-4-mercapto-N-benzyloxycarbonyl)pyrrolidine hydrochloride ##STR1472##

The same procedure as in Reference Example 42-1) was carried out by using (2S,4R)-2-(azetidin-3-yl)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxypyrrolidine (400 mg, 1.12 mmol) obtained by the reaction of Reference Example 41-12) to obtain (2S,4R)-N-tert-butoxycarbonyl-4-tert-butyldimethylsiloxy-2-(N-ethylazetidin-3-yl)pyrrolidine (280 mg, yield: 65%).

IR(KBr)cm⁻¹ : 1700, 1400, 1250, 1170 ##STR1473##

The same procedures as in Reference Example 38-6), 76), 8), 9), 10) were carried out by using the compound obtained by the above reaction (280 mg, 0.73 mmol) to obtain the above identified compound (113 mg).

IR(KBr)cm⁻¹ : 1700, 1520, 1350, 1110 

What is claimed as new and is desired to be secured by Letters Patent of the United States is:
 1. A compound of the formula: ##STR1474## wherein R is a hydrogen atom or a methyl group, R¹ is a hydrogen atom or a negative charge, each of R² and R³ which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, a formimidoyl group, an acetoimidoy 1 group, --COOR⁴, --CON(R⁵)R⁶, --N(R⁵)R⁶, --CH₂ COOR⁴, --CH₂ N(R⁵)R⁵ or --CH₂ CON(R⁵)R⁶ (wherein R⁴ is a hydrogen atom or a lower alkyl group, each of R⁵ and R⁶ which may be the same or different, is a hydrogen atom or a lower alkyl group, or R⁵ and R⁶ form together with the adjacent nitrogen atom a heterocyclic group selected from the group consisting of an aziridinyl group, an azetidinyl group, a pyrrolidinyl group and a piperidyl group) A is ═NR⁷, ═N(R⁷)R⁸, wherein each of R⁷ and R⁸ which may be the same or different, is a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, a formimidoyl group, an acetoimidoyl group, --COOR⁴, --CON(R⁵)R⁶), --N(R⁵)R⁶, --CH₂ COOR⁴, --CH₂ N(R⁵)R⁶ or --CH₂ CON(R⁵)R⁶ (wherein R⁴, R⁵ and R.sup. 6 are as defined above), p is an integer of from 0 to 3, and q is an integer of from 1 to 3; or a pharmaceutically acceptable salt or ester thereof.
 2. The compound according to claim 1, which is represented by the formula: ##STR1475## wherein R, R¹, R², R³, A, p, and q are as defined above; or a pharmaceutically acceptable salt or ester thereof.
 3. The compound according to claim 1, which is represented by the formula: ##STR1476## or a pharmaceutically acceptable salt or ester thereof.
 4. The compound according to claim 2, which is represented by the formula: ##STR1477## or a pharmaceutically acceptable salt or ester thereof. 